virology dr.titi
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SUNARYATI 1
Dept. Microbiology, Medical FacultyDept. Microbiology, Medical FacultyPadjadjaran UniversityPadjadjaran University
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SUNARYATI
a set of genesa set of genescomposed of either DNA or RNAcomposed of either DNA or RNApackages in a protein containing coatpackages in a protein containing coat
The resulting particle The resulting particle VirionVirion
Virus reproduction : virus particle infect a cell & Virus reproduction : virus particle infect a cell & program the cellular machinery to synthesize program the cellular machinery to synthesize the constituents required for the assembly of the constituents required for the assembly of new virions new virions considered an considered an intracellular intracellular parasiteparasite
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IntroductionIntroductionIntroductionIntroduction
Virus :Virus :
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3
Different viruses can have very different Different viruses can have very different genetics structures genetics structures reflected in their reflected in their replicative strategiesreplicative strategies
Because of their small size, viruses have Because of their small size, viruses have achieved a very high degree of genetic achieved a very high degree of genetic
economyeconomy
Viruses depend to great extent on host cell Viruses depend to great extent on host cell functions functions difficult to combat medically difficult to combat medically They do exhibit unique steps in their They do exhibit unique steps in their
replicative cycles that are potential targets replicative cycles that are potential targets for antiviral therapyfor antiviral therapy
Different viruses can have very different Different viruses can have very different genetics structures genetics structures reflected in their reflected in their replicative strategiesreplicative strategies
Because of their small size, viruses have Because of their small size, viruses have achieved a very high degree of genetic achieved a very high degree of genetic
economyeconomy
Viruses depend to great extent on host cell Viruses depend to great extent on host cell functions functions difficult to combat medically difficult to combat medically They do exhibit unique steps in their They do exhibit unique steps in their
replicative cycles that are potential targets replicative cycles that are potential targets for antiviral therapyfor antiviral therapy
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CHARACTERISTICCHARACTERISTICSS
CHARACTERISTICCHARACTERISTICSS
Small, ranging from about 20 – 300 nm diameter
Totally depend upon a living cell, either eukaryotic or prokaryotic, for replication and existence
Some viruses possess complex enzyme of their own : RNA or DNA polymerases but they cannot amplify & reproduce the information in their own genomes without assistance
Have a component a receptor-binding protein for attaching to cells
Small, ranging from about 20 – 300 nm diameter
Totally depend upon a living cell, either eukaryotic or prokaryotic, for replication and existence
Some viruses possess complex enzyme of their own : RNA or DNA polymerases but they cannot amplify & reproduce the information in their own genomes without assistance
Have a component a receptor-binding protein for attaching to cells
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Viral SizeViral Size
Length 20 - 14,000 nm
Much smaller than bacteria -
therefore filterable (infectious agent that could pass through porcelain filter; many originated in animals)
Length 20 - 14,000 nm
Much smaller than bacteria -
therefore filterable (infectious agent that could pass through porcelain filter; many originated in animals)
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SUNARYATI
CHARACTERISTICCHARACTERISTICSS
CHARACTERISTICCHARACTERISTICSS
To see the virus electron microscope
Growth need living cells/ tissues Can not growth saprophytic Only have certain enzyme for metabolism
and energy Easy mutated changes antigenic
property Multiplication different from bacteria
To see the virus electron microscope
Growth need living cells/ tissues Can not growth saprophytic Only have certain enzyme for metabolism
and energy Easy mutated changes antigenic
property Multiplication different from bacteria
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Virus structure & Virus structure & MorphologyMorphology
Virus structure & Virus structure & MorphologyMorphology
The basic design of all viruses places the The basic design of all viruses places the nucleic nucleic
acid genome on the inside of a protein shellacid genome on the inside of a protein shell capsid capsid
Two basic types of virions :Two basic types of virions :1. Enveloped viruses1. Enveloped viruses
have a nucleocapsid of nucleic acidhave a nucleocapsid of nucleic acid complexed to proteincomplexed to protein
2. Naked capsid viruses2. Naked capsid viruses have a nucleic acid genome within ahave a nucleic acid genome within a
protein shellprotein shell
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SUNARYATI 9
Virus structure & Virus structure & MorphologyMorphology
Virus structure & Virus structure & MorphologyMorphology
Schematic drawing of two basic type of virions
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Two basic shapes of virions :Two basic shapes of virions : 1. Cylindrical1. Cylindrical 2. Spherical2. Spherical
Some bacteriophages combine those 2 basic Some bacteriophages combine those 2 basic shapesshapes
Functions of capsid or envelope of viruses :Functions of capsid or envelope of viruses : 1. To protect the NA genome from damage 1. To protect the NA genome from damage during during extra-celullar passage of extra-celullar passage of the virus from the virus from one cell to anotherone cell to another 2. To aid in the process of entry into the cell2. To aid in the process of entry into the cell 3. To package enzymes essential for the early 3. To package enzymes essential for the early steps steps of the infection processof the infection process
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•
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Basic viral Basic viral formsforms
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The structure The structure and relative and relative
sizes of a sizes of a number of number of
DNADNA
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SUNARYATI 13
The structure The structure and relative and relative
sizes of a sizes of a number of number of
RNARNA
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DNA VIRUSESDNA VIRUSES
ENVELOPEDENVELOPED NAKED NAKED
Double – strandedDouble – stranded Double – stranded Single – Double – stranded Single – strandedstranded
Icosahedral Complex Icosahedral Icosahedral
HERPES POX PAPOVA PARVO
HEPADNA ADENO
ENVELOPEDENVELOPED NAKED NAKED
Double – strandedDouble – stranded Double – stranded Single – Double – stranded Single – strandedstranded
Icosahedral Complex Icosahedral Icosahedral
HERPES POX PAPOVA PARVO
HEPADNA ADENO
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RNA VIRUSESRNA VIRUSESRNA VIRUSESRNA VIRUSES Single – stranded Double – stranded
Positive – stranded (+) Negative – stranded (-)
Naked Enveloped Enveloped Naked
*PICORNA *TOGA BUNYA *REO*CALICI *FLAVI ORTHOMYXO
CORONA PARAMYXO RETRO RHABDO
ARENA FILO
* Icosahedral; all of the rest have helical symmetry
Single – stranded Double – stranded
Positive – stranded (+) Negative – stranded (-)
Naked Enveloped Enveloped Naked
*PICORNA *TOGA BUNYA *REO*CALICI *FLAVI ORTHOMYXO
CORONA PARAMYXO RETRO RHABDO
ARENA FILO
* Icosahedral; all of the rest have helical symmetry
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SUNARYATI
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SUNARYATI 18
The main groups of human viruses
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SUNARYATI 19
VIRAL REPLICATIONVIRAL REPLICATIONVIRAL REPLICATIONVIRAL REPLICATION
Virus multiplication cycle consist of 6 phases :
1. Attachment/ adsorption to the host cell2. Penetration or entry
3. Uncoating to release the genome4. Virion component production/ synthesis
5. Assembly/ maturation6. Release from the cells
Viral infections maybe productive/lytic response or nonproductive response (lysogeny)
Lysogeny may be associated with oncogenic transformation by animal viruses
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The outcome of the infection depend on :
the particular virus-host combination other factors : - the extracelullar environment - multiplicity of the infection - physiology & developmental state of the cell
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ADSORPTIONADSORPTIONADSORPTIONADSORPTIONAdsorption is the first step in every viral infection.
Adsorption involves :- virion attachment proteins- cell surface receptor proteins
Examples of viral receptors
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SUNARYATI 22
For some viruses co-receptors are involved in For some viruses co-receptors are involved in adsorption adsorption
HIV-1 : CD4 & chemokine receptorsHIV-1 : CD4 & chemokine receptors
Viral spikes & phage tails carry attachment proteins Viral spikes & phage tails carry attachment proteins
In some case, a region of the capsid protein serveIn some case, a region of the capsid protein serve the function of attachment the function of attachment
Adsorption is enhanced by presence of multiple Adsorption is enhanced by presence of multiple attachment & receptor proteins.attachment & receptor proteins.
A particular kind of virus is capable to infecting onlyA particular kind of virus is capable to infecting only a limited spectrum of cell types a limited spectrum of cell types its host rangeits host range Differences in host range & tissue tropism due Differences in host range & tissue tropism due
toto presence or absence of the receptors presence or absence of the receptors
For some viruses co-receptors are involved in For some viruses co-receptors are involved in adsorption adsorption
HIV-1 : CD4 & chemokine receptorsHIV-1 : CD4 & chemokine receptors
Viral spikes & phage tails carry attachment proteins Viral spikes & phage tails carry attachment proteins
In some case, a region of the capsid protein serveIn some case, a region of the capsid protein serve the function of attachment the function of attachment
Adsorption is enhanced by presence of multiple Adsorption is enhanced by presence of multiple attachment & receptor proteins.attachment & receptor proteins.
A particular kind of virus is capable to infecting onlyA particular kind of virus is capable to infecting only a limited spectrum of cell types a limited spectrum of cell types its host rangeits host range Differences in host range & tissue tropism due Differences in host range & tissue tropism due
toto presence or absence of the receptors presence or absence of the receptors
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SUNARYATI 23
Entry & UncoatingEntry & UncoatingEntry & UncoatingEntry & Uncoating
Enveloped Animal VirusesEnveloped Animal Viruses Some enveloped viruses enter cells by direct direct
fusionfusion of plasma membrane & envelope, release the nucleocapsid directly into the cytoplasm.
Paramyxoviruses (eg. measles) retroviruses (eg. HIV-1) & herpesviruses
Other enveloped & naked viruses are taken in by receptor-mediated endocytosisreceptor-mediated endocytosis (viropexis).
orthomycovirus (eg. influenza viruses), togaviruses (eg. rubella viruses), rhabdoviruses
(eg. rabies) & coronaviruses
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SUNARYATI 24
The two basic modes of entry of an enveloped animal virus
into the host cell
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Entry by Entry by
direct fusiondirect fusion
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SUNARYATI
ViropexisViropexis
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SUNARYATI 27
Naked Capsid Animal VirusesNaked Capsid Animal Viruses
Naked capsid viruses (eg. poliovirus, reovirus,Naked capsid viruses (eg. poliovirus, reovirus, adenovirus) also appear to enter by adenovirus) also appear to enter by viropexis, butviropexis, but in this case, virus can’t escape the in this case, virus can’t escape the endosomalendosomal vesicle by membrane fusion vesicle by membrane fusion
For poliovirus : acidified endosome exposeFor poliovirus : acidified endosome expose hydrophobic domains hydrophobic domains result in the binding result in the binding ofof the virions to membrane & releases the virions to membrane & releases nucleocapsidnucleocapsid to cytoplasm to cytoplasm
For reovirus : the content of the endosome areFor reovirus : the content of the endosome are transferred to a lysosome transferred to a lysosome the lysosomal the lysosomal
proteases strip away part of the capsid proteases strip away part of the capsid protein & protein & active virionsactive virions
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SUNARYATI 28Schematic of receptor-mediated endocytosis ulitilized by poliovirus for entry into host cell
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SUNARYATI 29
Insertion of Insertion of
glycoprotein glycoprotein
into the cell’s into the cell’s
membrane structures membrane structures
and formation of and formation of
the viral envelopethe viral envelope
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SUNARYATI 30
Viral release by Budding
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VIRAL LIFE
CYCLE
VIRAL LIFE
CYCLE
ATTACHMENT
PENETRATIONPENETRATION HOSTFUNCTIONS
ASSEMBLY(MATURATION)
Transcription
REPLICATION
RELEASE
UNCOATINGUNCOATING
Translation
MULTIPLICATION
Click after each step to view process
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Mechanisms of genetic changeMechanisms of genetic changeMechanisms of genetic changeMechanisms of genetic change
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MutationMutation
Many DNA viruses use the host DNA synthesis Many DNA viruses use the host DNA synthesis machinerymachinery
for replicating their genomes for replicating their genomes
However the largest animal viruses code for their However the largest animal viruses code for their own own DNA polymerases, & these enzymes are not DNA polymerases, & these enzymes are not as as effective at proofreading as the cellular effective at proofreading as the cellular polymerases polymerases
The resulting higher error rates in DNA The resulting higher error rates in DNA replication replication endow the viruses with the potential endow the viruses with the potential for a high rate for a high rate of evolution, but they are of evolution, but they are partially responsible for partially responsible for the high frequency of the high frequency of defective viral particles defective viral particles
High mutation rates permit adaptation to High mutation rates permit adaptation to changed changed
conditionsconditions
MutationMutation
Many DNA viruses use the host DNA synthesis Many DNA viruses use the host DNA synthesis machinerymachinery
for replicating their genomes for replicating their genomes
However the largest animal viruses code for their However the largest animal viruses code for their own own DNA polymerases, & these enzymes are not DNA polymerases, & these enzymes are not as as effective at proofreading as the cellular effective at proofreading as the cellular polymerases polymerases
The resulting higher error rates in DNA The resulting higher error rates in DNA replication replication endow the viruses with the potential endow the viruses with the potential for a high rate for a high rate of evolution, but they are of evolution, but they are partially responsible for partially responsible for the high frequency of the high frequency of defective viral particles defective viral particles
High mutation rates permit adaptation to High mutation rates permit adaptation to changed changed
conditionsconditions
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RecombinationRecombinationHomologous recombination is common in DNA Homologous recombination is common in DNA virusesviruses
There are 2 mechanisms : 1. Which is unique to the viruses with segmented genomes, involves reassortment of the segments during a mixed infections involving 2 different viral strains. 2. Exemplified by the genetic recombination between different forms of poliovirus Recombination occurs during replication by a “copy choice” type of mechanism Poliovirus replicase switches templates to generate recombinants 33
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COMMON METHODS OF INACTIVATING COMMON METHODS OF INACTIVATING VIRUSES FOR VARIOUS PURPOSEVIRUSES FOR VARIOUS PURPOSE
COMMON METHODS OF INACTIVATING COMMON METHODS OF INACTIVATING VIRUSES FOR VARIOUS PURPOSEVIRUSES FOR VARIOUS PURPOSE
Virus may be inactivated for : Sterilize laboratory supplies
Sterilization : steam, dry heat, ethylene oxide,
-irradiation Disinfect surfaces or skin
Surface disinfection : sodium hypochlorite, glutaraldehyde.
formaldehyde, peracetic acidSkin disinfection : chlorhexidine, 70% ethanol,
iodophores Vaccine production :
formaldehyde, -propiolactone, psoralen + uv irradiation, detergents (subunit vaccines)
Virus may be inactivated for : Sterilize laboratory supplies
Sterilization : steam, dry heat, ethylene oxide,
-irradiation Disinfect surfaces or skin
Surface disinfection : sodium hypochlorite, glutaraldehyde.
formaldehyde, peracetic acidSkin disinfection : chlorhexidine, 70% ethanol,
iodophores Vaccine production :
formaldehyde, -propiolactone, psoralen + uv irradiation, detergents (subunit vaccines)
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Cultivation in embryonated eggCultivation in embryonated egg
Virus injected into appropriate region
Method widely used for production of vaccines
Virus injected into appropriate region
Method widely used for production of vaccines
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Cultivation in cell cultures
Cultivation in cell cultures
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Cultivation in cell cultureCultivation in cell culture
Cytopathic effect - normal monolayer structure is disrupted by viral
infection Cell lines developed from embryonic
tissue Continuous cell lines (immortal) - HeLa Maintenance of cell culture lines is
technically difficult; must be kept free of microbial contamination.
Cytopathic effect - normal monolayer structure is disrupted by viral
infection Cell lines developed from embryonic
tissue Continuous cell lines (immortal) - HeLa Maintenance of cell culture lines is
technically difficult; must be kept free of microbial contamination.
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TRANSMISSION OF VIRAL INFECTIONTRANSMISSION OF VIRAL INFECTIONTRANSMISSION OF VIRAL INFECTIONTRANSMISSION OF VIRAL INFECTION
DIRECT CONTACTDIRECT CONTACTdroplet/aerosol
influenza virusmorbilli virussmallpox virus
GASTROINTESTINAL TRACT (ORALLY)GASTROINTESTINAL TRACT (ORALLY)enterovirusHepatitis A, E viruspoliomyelitis virus
ANIMAL BITEANIMAL BITE : rabies virus VECTORVECTOR : dengue virus PARENTERALPARENTERAL : Hepatitis B, C virus, HIV
DIRECT CONTACTDIRECT CONTACTdroplet/aerosol
influenza virusmorbilli virussmallpox virus
GASTROINTESTINAL TRACT (ORALLY)GASTROINTESTINAL TRACT (ORALLY)enterovirusHepatitis A, E viruspoliomyelitis virus
ANIMAL BITEANIMAL BITE : rabies virus VECTORVECTOR : dengue virus PARENTERALPARENTERAL : Hepatitis B, C virus, HIV
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PATHOGENESIS OF VIRAL DISEASESPATHOGENESIS OF VIRAL DISEASESPATHOGENESIS OF VIRAL DISEASESPATHOGENESIS OF VIRAL DISEASES
Viral pathogenesis :interaction of viral and host factors
leads to disease production
Virus pathogenic if : can infect and cause signs of disease of the host
Virus virulent : produce more severe disease Steps in viral pathogenesis :
Viral entry & primary replicationViral spread and cell tropismCell injury & clinical illnessRecovery from infectionVirus shedding
Viral pathogenesis :interaction of viral and host factors
leads to disease production
Virus pathogenic if : can infect and cause signs of disease of the host
Virus virulent : produce more severe disease Steps in viral pathogenesis :
Viral entry & primary replicationViral spread and cell tropismCell injury & clinical illnessRecovery from infectionVirus shedding
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DIAGNOSTIC METHODS OF VIRAL DIAGNOSTIC METHODS OF VIRAL INFECTIONINFECTION
DIAGNOSTIC METHODS OF VIRAL DIAGNOSTIC METHODS OF VIRAL INFECTIONINFECTION
1. Clinical symptoms2. Laboratory diagnosis
Typical clinical symptoms Polyomyelitis Chicken Pox Measles Mumps
1. Clinical symptoms2. Laboratory diagnosis
Typical clinical symptoms Polyomyelitis Chicken Pox Measles Mumps
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Laboratory Laboratory diagnosisdiagnosis
Laboratory Laboratory diagnosisdiagnosis
1. Electron microscope morphology2. Light microscope special staining
~ type of virus Variola virus inclusion bodies
Gispen staining : PASCHEN BODIES Rabies virus specimen : brain
Sellers staining : inclusion bodies in nerve cells NEGRI BODIES
Molluscum contagiosum virus skin nodule
Lugol staining : inclusion bodies in cytoplasm of epithel cell
MOLLUSCUM BODIES
1. Electron microscope morphology2. Light microscope special staining
~ type of virus Variola virus inclusion bodies
Gispen staining : PASCHEN BODIES Rabies virus specimen : brain
Sellers staining : inclusion bodies in nerve cells NEGRI BODIES
Molluscum contagiosum virus skin nodule
Lugol staining : inclusion bodies in cytoplasm of epithel cell
MOLLUSCUM BODIES
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Laboratory diagnosisLaboratory diagnosis Laboratory diagnosisLaboratory diagnosis
3. Culture specimen : depend on the diseasein vitro, in ovo, or in vivo
4. Serology Raise of antibody titer Antigen detection from the specimen Viral type identification :
agglutination, precipitation, complement fixation test, neutralization, inhibition haemagglutination, FAT, ELISA, RIA, LIA
3. Culture specimen : depend on the diseasein vitro, in ovo, or in vivo
4. Serology Raise of antibody titer Antigen detection from the specimen Viral type identification :
agglutination, precipitation, complement fixation test, neutralization, inhibition haemagglutination, FAT, ELISA, RIA, LIA
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PREVENTION AND TREATMENT OF VIRAL PREVENTION AND TREATMENT OF VIRAL INFECTIONSINFECTIONS
PREVENTION AND TREATMENT OF VIRAL PREVENTION AND TREATMENT OF VIRAL INFECTIONSINFECTIONS
1. VIRAL VACCINES Killed-virus vaccines Attenuated live-virus vaccines Future prospect : - attenuation of viruses by
genetic mapping - avirulent viral vectors - purified proteins produced using
cloned genes - synthetic peptides - subunit vaccines
- DNA vaccines
1. VIRAL VACCINES Killed-virus vaccines Attenuated live-virus vaccines Future prospect : - attenuation of viruses by
genetic mapping - avirulent viral vectors - purified proteins produced using
cloned genes - synthetic peptides - subunit vaccines
- DNA vaccines
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2. INTERFERONS2. INTERFERONS2. INTERFERONS2. INTERFERONS
IFNs : host-coded proteins of large
cytokine family inhibit viral replication produced by intact animal or
cell culture in response to viral infection or other inducers
first line of defense against viral infection
IFNs : host-coded proteins of large
cytokine family inhibit viral replication produced by intact animal or
cell culture in response to viral infection or other inducers
first line of defense against viral infection
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3. ANTIVIRAL CHEMOTHERAPY3. ANTIVIRAL CHEMOTHERAPY3. ANTIVIRAL CHEMOTHERAPY3. ANTIVIRAL CHEMOTHERAPY
A.A. Nucleoside Nucleoside analogsanalogs
Acyclovir & valacyclovir
Didanosine Gancyclovir Idoxuridine Lamivudin (3TC)
A.A. Nucleoside Nucleoside analogsanalogs
Acyclovir & valacyclovir
Didanosine Gancyclovir Idoxuridine Lamivudin (3TC)
Ribavirin Stavudine (d4T) Trifluridine Vidarabine Zalzitabine (ddC) Zidovudine (AZT)
Ribavirin Stavudine (d4T) Trifluridine Vidarabine Zalzitabine (ddC) Zidovudine (AZT)
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B. Nucleotide analogsB. Nucleotide analogs
Cidofovir : active against CMV & HSV inhibit s viral DNA
polymeraseC. Nonnucleoside reverse transcriptase C. Nonnucleoside reverse transcriptase
inhibitorinhibitorNevirapine : inhibit reverse transcriptase of Nevirapine : inhibit reverse transcriptase of HIV HIV
D. Protease inhibitorsRitonavir, Saquinavir HIV
E. Other types Amantadine & rimantadineFoscarnetMethiasone
Cidofovir : active against CMV & HSV inhibit s viral DNA
polymeraseC. Nonnucleoside reverse transcriptase C. Nonnucleoside reverse transcriptase
inhibitorinhibitorNevirapine : inhibit reverse transcriptase of Nevirapine : inhibit reverse transcriptase of HIV HIV
D. Protease inhibitorsRitonavir, Saquinavir HIV
E. Other types Amantadine & rimantadineFoscarnetMethiasone
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Questions ?Questions ?Questions ?Questions ?