Chapter 12Chapter 12

principles of principles of diagnosis and diagnosis and prevention of prevention of pathogen infectionpathogen infection

Bacteriological diagnosisBacteriological diagnosisVirology diagnosisVirology diagnosisMycology diagnosisMycology diagnosisSpecific prophylaxis and therapySpecific prophylaxis and therapy

section 1 bacterial diagnosissection 1 bacterial diagnosis

bacteriological diagnosisbacteriological diagnosischeck pathogen and their componentscheck pathogen and their components

（（ Ag products nucleic acidAg products nucleic acid ）） detect Ag with known Abmdash detect Ag with known Abmdash serological identificationserological identificationdetect Ab in serum detect Ab in serum detect unknown Ab with known Agmdashdetect unknown Ab with known Agmdash serodiagnosisserodiagnosis

Steps precede the laboratory Steps precede the laboratory workwork

11 Choose the appropriate specimenChoose the appropriate specimen

22 Obtain the specimen avoid contaminatedObtain the specimen avoid contaminated

33 Transport the specimen promptly or Transport the specimen promptly or store it correctlystore it correctly

44 Provide essential informationProvide essential information

（二）（二） detection of pathogenic bacteria detection of pathogenic bacteria

11 morphology and structuremorphology and structure

22 isolating cultureisolating culture

33 detection of Agdetection of Ag

44 other methodsother methods

11 morphosis detectionmorphosis detection

procedureprocedure smear fix stain observe under smear fix stain observe under

microscopemicroscope

（（ shapesizearrangement staining shapesizearrangement staining special structurespecial structure ） ） identify bacteriaidentify bacteria

major technique for detection of pathogmajor technique for detection of pathogenic bacteriaenic bacteria

（（ 11 ）） microscopemicroscope

Alight microscopeAlight microscope

resolutionresolution 025μm025μm

magnificationmagnification times1000times1000

B electronmicroscopeB electronmicroscope

resolutionresolution 1nm

magnification times100000

--cannot observe viable organism--cannot observe viable organism

C Other microscopeC Other microscope

bullbulldarkfield microscopedarkfield microscope

bull bullcontrast phase microscopecontrast phase microscope

bull bullfluorescence microscopefluorescence microscope

bull bullconfocal microscopeconfocal microscope

（（ 22 ）） staining techniquestaining technique

simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt

compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria

Gram stainingGram staining positivepositive （（ violetviolet ）） negativenegative （（ redred ））Acid-fast stainingAcid-fast staining positivepositive （（ redred ）） negative negative （（ blueblue ））Special stainingSpecial staining for bacterial capsule for bacterial capsule

spore flagella and so onspore flagella and so on

22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure

specimen

Plain agar

Blood agar

Selective medium

cultural feature

nutrition

growing condition

Colony feature

hemolysis

pure

inoculate

stain initial identify (morphology)

Biochemical reaction

Serological identificationSerological identification species groupspecies groupss

Virulence Virulence identificationidentification

detect endotoxin or exotoxindetect endotoxin or exotoxin

animal testanimal test rabbit guinea pirabbit guinea pigg

Susceptibility Susceptibility testtest

Slip methodSlip method determine the naturedetermine the nature

Test tube methodTest tube method quantitationquantitation（（ detect MBCdetect MBC MICMIC ））

33 detect Agdetect Ag

mdashmdash mdashmdashserological identificationserological identification

highly specificityhighly specificity

sensitivity sensitivity

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

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• Slide 49
• Slide 50

Bacteriological diagnosisBacteriological diagnosisVirology diagnosisVirology diagnosisMycology diagnosisMycology diagnosisSpecific prophylaxis and therapySpecific prophylaxis and therapy

section 1 bacterial diagnosissection 1 bacterial diagnosis

bacteriological diagnosisbacteriological diagnosischeck pathogen and their componentscheck pathogen and their components

（（ Ag products nucleic acidAg products nucleic acid ）） detect Ag with known Abmdash detect Ag with known Abmdash serological identificationserological identificationdetect Ab in serum detect Ab in serum detect unknown Ab with known Agmdashdetect unknown Ab with known Agmdash serodiagnosisserodiagnosis

Steps precede the laboratory Steps precede the laboratory workwork

11 Choose the appropriate specimenChoose the appropriate specimen

22 Obtain the specimen avoid contaminatedObtain the specimen avoid contaminated

33 Transport the specimen promptly or Transport the specimen promptly or store it correctlystore it correctly

44 Provide essential informationProvide essential information

（二）（二） detection of pathogenic bacteria detection of pathogenic bacteria

11 morphology and structuremorphology and structure

22 isolating cultureisolating culture

33 detection of Agdetection of Ag

44 other methodsother methods

11 morphosis detectionmorphosis detection

procedureprocedure smear fix stain observe under smear fix stain observe under

microscopemicroscope

（（ shapesizearrangement staining shapesizearrangement staining special structurespecial structure ） ） identify bacteriaidentify bacteria

major technique for detection of pathogmajor technique for detection of pathogenic bacteriaenic bacteria

（（ 11 ）） microscopemicroscope

Alight microscopeAlight microscope

resolutionresolution 025μm025μm

magnificationmagnification times1000times1000

B electronmicroscopeB electronmicroscope

resolutionresolution 1nm

magnification times100000

--cannot observe viable organism--cannot observe viable organism

C Other microscopeC Other microscope

bullbulldarkfield microscopedarkfield microscope

bull bullcontrast phase microscopecontrast phase microscope

bull bullfluorescence microscopefluorescence microscope

bull bullconfocal microscopeconfocal microscope

（（ 22 ）） staining techniquestaining technique

simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt

compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria

Gram stainingGram staining positivepositive （（ violetviolet ）） negativenegative （（ redred ））Acid-fast stainingAcid-fast staining positivepositive （（ redred ）） negative negative （（ blueblue ））Special stainingSpecial staining for bacterial capsule for bacterial capsule

spore flagella and so onspore flagella and so on

22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure

specimen

Plain agar

Blood agar

Selective medium

cultural feature

nutrition

growing condition

Colony feature

hemolysis

pure

inoculate

stain initial identify (morphology)

Biochemical reaction

Serological identificationSerological identification species groupspecies groupss

Virulence Virulence identificationidentification

detect endotoxin or exotoxindetect endotoxin or exotoxin

animal testanimal test rabbit guinea pirabbit guinea pigg

Susceptibility Susceptibility testtest

Slip methodSlip method determine the naturedetermine the nature

Test tube methodTest tube method quantitationquantitation（（ detect MBCdetect MBC MICMIC ））

33 detect Agdetect Ag

mdashmdash mdashmdashserological identificationserological identification

highly specificityhighly specificity

sensitivity sensitivity

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
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• Slide 48
• Slide 49
• Slide 50

section 1 bacterial diagnosissection 1 bacterial diagnosis

bacteriological diagnosisbacteriological diagnosischeck pathogen and their componentscheck pathogen and their components

（（ Ag products nucleic acidAg products nucleic acid ）） detect Ag with known Abmdash detect Ag with known Abmdash serological identificationserological identificationdetect Ab in serum detect Ab in serum detect unknown Ab with known Agmdashdetect unknown Ab with known Agmdash serodiagnosisserodiagnosis

Steps precede the laboratory Steps precede the laboratory workwork

11 Choose the appropriate specimenChoose the appropriate specimen

22 Obtain the specimen avoid contaminatedObtain the specimen avoid contaminated

33 Transport the specimen promptly or Transport the specimen promptly or store it correctlystore it correctly

44 Provide essential informationProvide essential information

（二）（二） detection of pathogenic bacteria detection of pathogenic bacteria

11 morphology and structuremorphology and structure

22 isolating cultureisolating culture

33 detection of Agdetection of Ag

44 other methodsother methods

11 morphosis detectionmorphosis detection

procedureprocedure smear fix stain observe under smear fix stain observe under

microscopemicroscope

（（ shapesizearrangement staining shapesizearrangement staining special structurespecial structure ） ） identify bacteriaidentify bacteria

major technique for detection of pathogmajor technique for detection of pathogenic bacteriaenic bacteria

（（ 11 ）） microscopemicroscope

Alight microscopeAlight microscope

resolutionresolution 025μm025μm

magnificationmagnification times1000times1000

B electronmicroscopeB electronmicroscope

resolutionresolution 1nm

magnification times100000

--cannot observe viable organism--cannot observe viable organism

C Other microscopeC Other microscope

bullbulldarkfield microscopedarkfield microscope

bull bullcontrast phase microscopecontrast phase microscope

bull bullfluorescence microscopefluorescence microscope

bull bullconfocal microscopeconfocal microscope

（（ 22 ）） staining techniquestaining technique

simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt

compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria

Gram stainingGram staining positivepositive （（ violetviolet ）） negativenegative （（ redred ））Acid-fast stainingAcid-fast staining positivepositive （（ redred ）） negative negative （（ blueblue ））Special stainingSpecial staining for bacterial capsule for bacterial capsule

spore flagella and so onspore flagella and so on

22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure

specimen

Plain agar

Blood agar

Selective medium

cultural feature

nutrition

growing condition

Colony feature

hemolysis

pure

inoculate

stain initial identify (morphology)

Biochemical reaction

Serological identificationSerological identification species groupspecies groupss

Virulence Virulence identificationidentification

detect endotoxin or exotoxindetect endotoxin or exotoxin

animal testanimal test rabbit guinea pirabbit guinea pigg

Susceptibility Susceptibility testtest

Slip methodSlip method determine the naturedetermine the nature

Test tube methodTest tube method quantitationquantitation（（ detect MBCdetect MBC MICMIC ））

33 detect Agdetect Ag

mdashmdash mdashmdashserological identificationserological identification

highly specificityhighly specificity

sensitivity sensitivity

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
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• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Slide 18
• Slide 19
• Slide 20
• Slide 21
• Slide 22
• Slide 23
• Slide 24
• Slide 25
• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

Steps precede the laboratory Steps precede the laboratory workwork

11 Choose the appropriate specimenChoose the appropriate specimen

22 Obtain the specimen avoid contaminatedObtain the specimen avoid contaminated

33 Transport the specimen promptly or Transport the specimen promptly or store it correctlystore it correctly

44 Provide essential informationProvide essential information

（二）（二） detection of pathogenic bacteria detection of pathogenic bacteria

11 morphology and structuremorphology and structure

22 isolating cultureisolating culture

33 detection of Agdetection of Ag

44 other methodsother methods

11 morphosis detectionmorphosis detection

procedureprocedure smear fix stain observe under smear fix stain observe under

microscopemicroscope

（（ shapesizearrangement staining shapesizearrangement staining special structurespecial structure ） ） identify bacteriaidentify bacteria

major technique for detection of pathogmajor technique for detection of pathogenic bacteriaenic bacteria

（（ 11 ）） microscopemicroscope

Alight microscopeAlight microscope

resolutionresolution 025μm025μm

magnificationmagnification times1000times1000

B electronmicroscopeB electronmicroscope

resolutionresolution 1nm

magnification times100000

--cannot observe viable organism--cannot observe viable organism

C Other microscopeC Other microscope

bullbulldarkfield microscopedarkfield microscope

bull bullcontrast phase microscopecontrast phase microscope

bull bullfluorescence microscopefluorescence microscope

bull bullconfocal microscopeconfocal microscope

（（ 22 ）） staining techniquestaining technique

simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt

compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria

Gram stainingGram staining positivepositive （（ violetviolet ）） negativenegative （（ redred ））Acid-fast stainingAcid-fast staining positivepositive （（ redred ）） negative negative （（ blueblue ））Special stainingSpecial staining for bacterial capsule for bacterial capsule

spore flagella and so onspore flagella and so on

22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure

specimen

Plain agar

Blood agar

Selective medium

cultural feature

nutrition

growing condition

Colony feature

hemolysis

pure

inoculate

stain initial identify (morphology)

Biochemical reaction

Serological identificationSerological identification species groupspecies groupss

Virulence Virulence identificationidentification

detect endotoxin or exotoxindetect endotoxin or exotoxin

animal testanimal test rabbit guinea pirabbit guinea pigg

Susceptibility Susceptibility testtest

Slip methodSlip method determine the naturedetermine the nature

Test tube methodTest tube method quantitationquantitation（（ detect MBCdetect MBC MICMIC ））

33 detect Agdetect Ag

mdashmdash mdashmdashserological identificationserological identification

highly specificityhighly specificity

sensitivity sensitivity

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
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• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

（二）（二） detection of pathogenic bacteria detection of pathogenic bacteria

11 morphology and structuremorphology and structure

22 isolating cultureisolating culture

33 detection of Agdetection of Ag

44 other methodsother methods

11 morphosis detectionmorphosis detection

procedureprocedure smear fix stain observe under smear fix stain observe under

microscopemicroscope

（（ shapesizearrangement staining shapesizearrangement staining special structurespecial structure ） ） identify bacteriaidentify bacteria

major technique for detection of pathogmajor technique for detection of pathogenic bacteriaenic bacteria

（（ 11 ）） microscopemicroscope

Alight microscopeAlight microscope

resolutionresolution 025μm025μm

magnificationmagnification times1000times1000

B electronmicroscopeB electronmicroscope

resolutionresolution 1nm

magnification times100000

--cannot observe viable organism--cannot observe viable organism

C Other microscopeC Other microscope

bullbulldarkfield microscopedarkfield microscope

bull bullcontrast phase microscopecontrast phase microscope

bull bullfluorescence microscopefluorescence microscope

bull bullconfocal microscopeconfocal microscope

（（ 22 ）） staining techniquestaining technique

simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt

compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria

Gram stainingGram staining positivepositive （（ violetviolet ）） negativenegative （（ redred ））Acid-fast stainingAcid-fast staining positivepositive （（ redred ）） negative negative （（ blueblue ））Special stainingSpecial staining for bacterial capsule for bacterial capsule

spore flagella and so onspore flagella and so on

22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure

specimen

Plain agar

Blood agar

Selective medium

cultural feature

nutrition

growing condition

Colony feature

hemolysis

pure

inoculate

stain initial identify (morphology)

Biochemical reaction

Serological identificationSerological identification species groupspecies groupss

Virulence Virulence identificationidentification

detect endotoxin or exotoxindetect endotoxin or exotoxin

animal testanimal test rabbit guinea pirabbit guinea pigg

Susceptibility Susceptibility testtest

Slip methodSlip method determine the naturedetermine the nature

Test tube methodTest tube method quantitationquantitation（（ detect MBCdetect MBC MICMIC ））

33 detect Agdetect Ag

mdashmdash mdashmdashserological identificationserological identification

highly specificityhighly specificity

sensitivity sensitivity

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Slide 18
• Slide 19
• Slide 20
• Slide 21
• Slide 22
• Slide 23
• Slide 24
• Slide 25
• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

11 morphosis detectionmorphosis detection

procedureprocedure smear fix stain observe under smear fix stain observe under

microscopemicroscope

（（ shapesizearrangement staining shapesizearrangement staining special structurespecial structure ） ） identify bacteriaidentify bacteria

major technique for detection of pathogmajor technique for detection of pathogenic bacteriaenic bacteria

（（ 11 ）） microscopemicroscope

Alight microscopeAlight microscope

resolutionresolution 025μm025μm

magnificationmagnification times1000times1000

B electronmicroscopeB electronmicroscope

resolutionresolution 1nm

magnification times100000

--cannot observe viable organism--cannot observe viable organism

C Other microscopeC Other microscope

bullbulldarkfield microscopedarkfield microscope

bull bullcontrast phase microscopecontrast phase microscope

bull bullfluorescence microscopefluorescence microscope

bull bullconfocal microscopeconfocal microscope

（（ 22 ）） staining techniquestaining technique

simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt

compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria

Gram stainingGram staining positivepositive （（ violetviolet ）） negativenegative （（ redred ））Acid-fast stainingAcid-fast staining positivepositive （（ redred ）） negative negative （（ blueblue ））Special stainingSpecial staining for bacterial capsule for bacterial capsule

spore flagella and so onspore flagella and so on

22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure

specimen

Plain agar

Blood agar

Selective medium

cultural feature

nutrition

growing condition

Colony feature

hemolysis

pure

inoculate

stain initial identify (morphology)

Biochemical reaction

Serological identificationSerological identification species groupspecies groupss

Virulence Virulence identificationidentification

detect endotoxin or exotoxindetect endotoxin or exotoxin

animal testanimal test rabbit guinea pirabbit guinea pigg

Susceptibility Susceptibility testtest

Slip methodSlip method determine the naturedetermine the nature

Test tube methodTest tube method quantitationquantitation（（ detect MBCdetect MBC MICMIC ））

33 detect Agdetect Ag

mdashmdash mdashmdashserological identificationserological identification

highly specificityhighly specificity

sensitivity sensitivity

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Slide 18
• Slide 19
• Slide 20
• Slide 21
• Slide 22
• Slide 23
• Slide 24
• Slide 25
• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

major technique for detection of pathogmajor technique for detection of pathogenic bacteriaenic bacteria

（（ 11 ）） microscopemicroscope

Alight microscopeAlight microscope

resolutionresolution 025μm025μm

magnificationmagnification times1000times1000

B electronmicroscopeB electronmicroscope

resolutionresolution 1nm

magnification times100000

--cannot observe viable organism--cannot observe viable organism

C Other microscopeC Other microscope

bullbulldarkfield microscopedarkfield microscope

bull bullcontrast phase microscopecontrast phase microscope

bull bullfluorescence microscopefluorescence microscope

bull bullconfocal microscopeconfocal microscope

（（ 22 ）） staining techniquestaining technique

simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt

compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria

Gram stainingGram staining positivepositive （（ violetviolet ）） negativenegative （（ redred ））Acid-fast stainingAcid-fast staining positivepositive （（ redred ）） negative negative （（ blueblue ））Special stainingSpecial staining for bacterial capsule for bacterial capsule

spore flagella and so onspore flagella and so on

22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure

specimen

Plain agar

Blood agar

Selective medium

cultural feature

nutrition

growing condition

Colony feature

hemolysis

pure

inoculate

stain initial identify (morphology)

Biochemical reaction

Serological identificationSerological identification species groupspecies groupss

Virulence Virulence identificationidentification

detect endotoxin or exotoxindetect endotoxin or exotoxin

animal testanimal test rabbit guinea pirabbit guinea pigg

Susceptibility Susceptibility testtest

Slip methodSlip method determine the naturedetermine the nature

Test tube methodTest tube method quantitationquantitation（（ detect MBCdetect MBC MICMIC ））

33 detect Agdetect Ag

mdashmdash mdashmdashserological identificationserological identification

highly specificityhighly specificity

sensitivity sensitivity

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Slide 18
• Slide 19
• Slide 20
• Slide 21
• Slide 22
• Slide 23
• Slide 24
• Slide 25
• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

B electronmicroscopeB electronmicroscope

resolutionresolution 1nm

magnification times100000

--cannot observe viable organism--cannot observe viable organism

C Other microscopeC Other microscope

bullbulldarkfield microscopedarkfield microscope

bull bullcontrast phase microscopecontrast phase microscope

bull bullfluorescence microscopefluorescence microscope

bull bullconfocal microscopeconfocal microscope

（（ 22 ）） staining techniquestaining technique

simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt

compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria

Gram stainingGram staining positivepositive （（ violetviolet ）） negativenegative （（ redred ））Acid-fast stainingAcid-fast staining positivepositive （（ redred ）） negative negative （（ blueblue ））Special stainingSpecial staining for bacterial capsule for bacterial capsule

spore flagella and so onspore flagella and so on

22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure

specimen

Plain agar

Blood agar

Selective medium

cultural feature

nutrition

growing condition

Colony feature

hemolysis

pure

inoculate

stain initial identify (morphology)

Biochemical reaction

Serological identificationSerological identification species groupspecies groupss

Virulence Virulence identificationidentification

detect endotoxin or exotoxindetect endotoxin or exotoxin

animal testanimal test rabbit guinea pirabbit guinea pigg

Susceptibility Susceptibility testtest

Slip methodSlip method determine the naturedetermine the nature

Test tube methodTest tube method quantitationquantitation（（ detect MBCdetect MBC MICMIC ））

33 detect Agdetect Ag

mdashmdash mdashmdashserological identificationserological identification

highly specificityhighly specificity

sensitivity sensitivity

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Slide 18
• Slide 19
• Slide 20
• Slide 21
• Slide 22
• Slide 23
• Slide 24
• Slide 25
• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

C Other microscopeC Other microscope

bullbulldarkfield microscopedarkfield microscope

bull bullcontrast phase microscopecontrast phase microscope

bull bullfluorescence microscopefluorescence microscope

bull bullconfocal microscopeconfocal microscope

（（ 22 ）） staining techniquestaining technique

simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt

compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria

Gram stainingGram staining positivepositive （（ violetviolet ）） negativenegative （（ redred ））Acid-fast stainingAcid-fast staining positivepositive （（ redred ）） negative negative （（ blueblue ））Special stainingSpecial staining for bacterial capsule for bacterial capsule

spore flagella and so onspore flagella and so on

22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure

specimen

Plain agar

Blood agar

Selective medium

cultural feature

nutrition

growing condition

Colony feature

hemolysis

pure

inoculate

stain initial identify (morphology)

Biochemical reaction

Serological identificationSerological identification species groupspecies groupss

Virulence Virulence identificationidentification

detect endotoxin or exotoxindetect endotoxin or exotoxin

animal testanimal test rabbit guinea pirabbit guinea pigg

Susceptibility Susceptibility testtest

Slip methodSlip method determine the naturedetermine the nature

Test tube methodTest tube method quantitationquantitation（（ detect MBCdetect MBC MICMIC ））

33 detect Agdetect Ag

mdashmdash mdashmdashserological identificationserological identification

highly specificityhighly specificity

sensitivity sensitivity

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Slide 18
• Slide 19
• Slide 20
• Slide 21
• Slide 22
• Slide 23
• Slide 24
• Slide 25
• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

（（ 22 ）） staining techniquestaining technique

simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt

compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria

Gram stainingGram staining positivepositive （（ violetviolet ）） negativenegative （（ redred ））Acid-fast stainingAcid-fast staining positivepositive （（ redred ）） negative negative （（ blueblue ））Special stainingSpecial staining for bacterial capsule for bacterial capsule

spore flagella and so onspore flagella and so on

22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure

specimen

Plain agar

Blood agar

Selective medium

cultural feature

nutrition

growing condition

Colony feature

hemolysis

pure

inoculate

stain initial identify (morphology)

Biochemical reaction

Serological identificationSerological identification species groupspecies groupss

Virulence Virulence identificationidentification

detect endotoxin or exotoxindetect endotoxin or exotoxin

animal testanimal test rabbit guinea pirabbit guinea pigg

Susceptibility Susceptibility testtest

Slip methodSlip method determine the naturedetermine the nature

Test tube methodTest tube method quantitationquantitation（（ detect MBCdetect MBC MICMIC ））

33 detect Agdetect Ag

mdashmdash mdashmdashserological identificationserological identification

highly specificityhighly specificity

sensitivity sensitivity

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Slide 18
• Slide 19
• Slide 20
• Slide 21
• Slide 22
• Slide 23
• Slide 24
• Slide 25
• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

Gram stainingGram staining positivepositive （（ violetviolet ）） negativenegative （（ redred ））Acid-fast stainingAcid-fast staining positivepositive （（ redred ）） negative negative （（ blueblue ））Special stainingSpecial staining for bacterial capsule for bacterial capsule

spore flagella and so onspore flagella and so on

22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure

specimen

Plain agar

Blood agar

Selective medium

cultural feature

nutrition

growing condition

Colony feature

hemolysis

pure

inoculate

stain initial identify (morphology)

Biochemical reaction

Serological identificationSerological identification species groupspecies groupss

Virulence Virulence identificationidentification

detect endotoxin or exotoxindetect endotoxin or exotoxin

animal testanimal test rabbit guinea pirabbit guinea pigg

Susceptibility Susceptibility testtest

Slip methodSlip method determine the naturedetermine the nature

Test tube methodTest tube method quantitationquantitation（（ detect MBCdetect MBC MICMIC ））

33 detect Agdetect Ag

mdashmdash mdashmdashserological identificationserological identification

highly specificityhighly specificity

sensitivity sensitivity

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Slide 18
• Slide 19
• Slide 20
• Slide 21
• Slide 22
• Slide 23
• Slide 24
• Slide 25
• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure

specimen

Plain agar

Blood agar

Selective medium

cultural feature

nutrition

growing condition

Colony feature

hemolysis

pure

inoculate

stain initial identify (morphology)

Biochemical reaction

Serological identificationSerological identification species groupspecies groupss

Virulence Virulence identificationidentification

detect endotoxin or exotoxindetect endotoxin or exotoxin

animal testanimal test rabbit guinea pirabbit guinea pigg

Susceptibility Susceptibility testtest

Slip methodSlip method determine the naturedetermine the nature

Test tube methodTest tube method quantitationquantitation（（ detect MBCdetect MBC MICMIC ））

33 detect Agdetect Ag

mdashmdash mdashmdashserological identificationserological identification

highly specificityhighly specificity

sensitivity sensitivity

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
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• Slide 14
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• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

Serological identificationSerological identification species groupspecies groupss

Virulence Virulence identificationidentification

detect endotoxin or exotoxindetect endotoxin or exotoxin

animal testanimal test rabbit guinea pirabbit guinea pigg

Susceptibility Susceptibility testtest

Slip methodSlip method determine the naturedetermine the nature

Test tube methodTest tube method quantitationquantitation（（ detect MBCdetect MBC MICMIC ））

33 detect Agdetect Ag

mdashmdash mdashmdashserological identificationserological identification

highly specificityhighly specificity

sensitivity sensitivity

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Slide 18
• Slide 19
• Slide 20
• Slide 21
• Slide 22
• Slide 23
• Slide 24
• Slide 25
• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

33 detect Agdetect Ag

mdashmdash mdashmdashserological identificationserological identification

highly specificityhighly specificity

sensitivity sensitivity

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Slide 18
• Slide 19
• Slide 20
• Slide 21
• Slide 22
• Slide 23
• Slide 24
• Slide 25
• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

44 other detectionother detection

Detect metabolic productDetect metabolic product

bull bull PCR detect DNAPCR detect DNA

caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)

bull bull DNA chips techniqueDNA chips technique

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
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• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

22 serological diagnosisserological diagnosis

-----detect Ab with known Ag-----detect Ab with known Ag

------specimens------specimens serumserum

------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Slide 18
• Slide 19
• Slide 20
• Slide 21
• Slide 22
• Slide 23
• Slide 24
• Slide 25
• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively

A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection

Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
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• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Slide 18
• Slide 19
• Slide 20
• Slide 21
• Slide 22
• Slide 23
• Slide 24
• Slide 25
• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
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• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

commonly used methodscommonly used methods

Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and

paratyphoidparatyphoid

Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis

10486801048680

Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic

diseasedisease

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
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• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

Section 2 virology diagnosisSection 2 virology diagnosis

ViralViral infeinfectionction

Electron microscopeElectron microscope

Light microscope inLight microscope inclusion bodyclusion body

isolationisolation

VirusVirus particleparticle

V AgV Ag

V nucleic acidV nucleic acid

Viral enzymeViral enzyme retroviridaseretroviridase

DetectDetect virusvirus

DetectDetect AbAb

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
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• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

Choose the appropriate specimenChoose the appropriate specimen

Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci

menmen

Take it in acute phase Take it in acute phase

Low temperature conservationLow temperature conservation transport it transport it

promptlypromptly preserve in 50preserve in 50 glycerineglycerine

Serologial diagnose paired serumSerologial diagnose paired serum

Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Slide 18
• Slide 19
• Slide 20
• Slide 21
• Slide 22
• Slide 23
• Slide 24
• Slide 25
• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

二二 viral isolation and identifyingviral isolation and identifying

specimenspecimenAnimal inoculaAnimal inoculationtion

Embryonated eEmbryonated egggg

Cell cultureCell culture

positivepositive

negativenegative

identifyingidentifying

BlindBlind passagepassage22 － 3generatiogenerationn

negativenegative

Virus negatiVirus negativeve

asepsisasepsis

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
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• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

（一）（一） viral isolationviral isolation

11 animal inoculationanimal inoculation

chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
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• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

22 embryonated egg inoculationembryonated egg inoculation

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
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• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

3 Cell culture3 Cell culture （（ commonly usedcommonly used ））Primary and secondary cell culturePrimary and secondary cell culture

general proceduregeneral procedure

tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell

cell monolayercell monolayer

brokenbroken proteaseprotease

(primary cell)(primary cell)Serial subcultivationSerial subcultivation

Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
• Slide 12
• Slide 13
• Slide 14
• Slide 15
• Slide 16
• Slide 17
• Slide 18
• Slide 19
• Slide 20
• Slide 21
• Slide 22
• Slide 23
• Slide 24
• Slide 25
• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

features of virus cell culturefeatures of virus cell culture

A sensitive to many kinds of virusesA sensitive to many kinds of viruses

B cost highB cost high

C carry latent virusC carry latent virus

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
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• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

(2)Diploid(2)Diploid cellcell cultureculture

usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
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• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

(3) secondary cell culture(3) secondary cell culture

sensitive to sensitive to multiplemultiple viruses viruses

high reproductive capacity long genera high reproductive capacity long generation timetion time

have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
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• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

(( 二二 )) indexindex for reproduction of virusfor reproduction of virus

(1)Virus1)Virus infected cellinfected cell

（（ cell roundinggathercell roundinggather

necrosisnecrosis ））

1 Cytopathic effect CPE)1 Cytopathic effect CPE)

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
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• Slide 33
• Slide 34
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• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

CPECPE

（（ 22 ）） cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
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• Slide 22
• Slide 23
• Slide 24
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• Slide 26
• Slide 27
• Slide 28
• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

（（ 33 ）） viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell

CPECPE

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
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• Slide 47
• Slide 48
• Slide 49
• Slide 50

22 hemadsorption hemadsorption HAd HAd

hemagglutininhemagglutinin （（ HAHA ）） on membrane of vion membrane of virus infected cell rus infected cell

Can Can adsorbadsorb vertebrate vertebrate RBCRBC

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
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• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

3 Interference3 Interference

4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
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• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

( 三 ) viral quantitative assayHemagglutination test total amount

Plaque bacteriophage determination

ID50 or TCID50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
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• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le

sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex

cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a

nd hyphand hypha

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
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• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

22 cultureculture

specimen sabouraud mediumspecimen sabouraud medium

macroculture microculturemacroculture microculture

colonycolony

biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique

sporespore

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
• Slide 2
• Slide 3
• Slide 4
• Slide 5
• Slide 6
• Slide 7
• Slide 8
• Slide 9
• Slide 10
• Slide 11
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• Slide 17
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• Slide 26
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• Slide 29
• Slide 30
• Slide 31
• Slide 32
• Slide 33
• Slide 34
• Slide 35
• Slide 36
• Slide 37
• Slide 39
• Slide 40
• Slide 41
• Slide 42
• Slide 43
• Slide 44
• Slide 45
• Slide 46
• Slide 47
• Slide 48
• Slide 49
• Slide 50

3animal test3animal test

4skin test hypersensitivy4skin test hypersensitivy

（三）（三） mycology rapid diagnosismycology rapid diagnosis

serological test detect Ag or Abserological test detect Ag or Ab

detect nucleic acid of fungusdetect nucleic acid of fungus

detect mycotoxindetect mycotoxin

questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

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questionsquestions

11 definitiondefinition

serological identificationserological identification

and diagnosis)and diagnosis)

Cytopathic effect CPECytopathic effect CPE

hemadsorption HAdhemadsorption HAd

interferenceinterference

22 principle of Widalrsquos testprinciple of Widalrsquos test

33 principle of Gramrsquos stainingprinciple of Gramrsquos staining

Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

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Section 4Section 4

artificial active artificial active immunityimmunity

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

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• Slide 50

Artificial active immunityArtificial active immunity

stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid

vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

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vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA

A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

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A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow

th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni

citycity are repeatedly selected are repeatedly selected BCGBCG

The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

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The mutation is expressed only The mutation is expressed only

under certain conditionsunder certain conditions

may be may be useful in vaccinesuseful in vaccines

Conditional-lethal mutationsConditional-lethal mutations

temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

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temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation

the organisms can the organisms can replicate replicate at a at a

relativelyrelatively low low permissive temperature permissive temperature

but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive

temperaturetemperature

Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
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Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize

d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost

errdquo injectionerrdquo injection

A Live (attenuated) A Live (attenuated) vaccinesvaccines

B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

• Slide 1
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B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate

responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe

toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

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toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves

artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

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artificial passive artificial passive immunization immunization

Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

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Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac

tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors

who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized

Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

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Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals

Cannot form AbsCannot form Abs Before Ab production Before Ab production

animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions

Artificial passive Artificial passive immunizationimmunization

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