06 microbiology - specimen handling collection instructions

8/22/2019 06 Microbiology - Specimen Handling Collection Instructions

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MICROBIOLOGY

SPECIMEN HANDL ING & COL LECTION INSTRUCTIONS

Doc#8231 Ver.#3.0 Date: 15-Apr-13 Page 1 of 21

For Inquires, contact LifeLabs Customer Care Centre - Toll Free: 877-849-3637 or GTA: 416-675-3637Printed Copies are uncontrolled

TABL E OF CONTENTS Page

I. Collection of Specimens for Microbiolog y 3II. Microb iol ogy Swab and Collectio n Containers 4-5

III. Urine for Culture

Collection of Midstream Urine Samples

Urine for TB Culture

6

IV. Swabs fo r Culture

Collection of Wound Swabs

Collection of Eye Swabs

Collection of Ear Swabs

Collection of Throat Swabs

Collection of Urethral Swabs for Culture for Neisseria gonorrhoeae

Collection of Endocervical Swabs for Culture ofNeisseria gonorrhoeae

Collection of Vaginal Swabs

Collection of Vaginal/Rectal Swabs for Prenatal Group B StreptococcusScreening at 35-37 Weeks Gestation

7-9

V. Chlamydia trachomatis : Nucleic Acid Amplif ication Testing &

Neisseria gonorrhoeae: Nucleic Acid Amplif i cation Testing

Collection of Endocervical Swab for CT/N. gonorrhoeae NAAT

Collection of Urethral Swab for CT/N. gonorrhoeae NAATCollection of Urine for CT/N. gonorrhoeae NAAT

10-11

VI. Chlamydia Culture

Chlamydia Culture- Non-genital Swabs (rectal, eye, conjunctivialscrapings and swabs, pharyngeal)

12

VII. Surveillance Specimens 12

VIII. Stool Specimens

Collection of Stool Specimens

Stool Culture

Ova and Parasites

Clostridium difficile Toxin

13-14

IX. Parasitology Specimens

Collection for Pinworm Examination

Collection of Urine for Schistosoma

Collection of Specimens for Identification (non-stool): Worms, Insect,Proglottid, Skin Scrapings for Scabies, Hair Specimens for Lice

14


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For Inquires, contact LifeLabs Customer Care Centre - Toll Free: 877-849-3637 or GTA: 416-675-3637

Printed Copies are uncontrolled

TABLE OF CONTENTS Page

X. Fungus/Mycol ogy for Dermatophyt es

Nail, Skin and Hair SpecimensVaginal, Throat, Mouth and Ear SwabsCollection of Nail Specimen

15

XI. Sterile Flui ds (Other than Blood & CSF) 16

XII. Bloo d Cultures

Adult CollectionChild CollectionRequest for Fungus, Yeast, Cryptococcus, Mycobacteria, TB, AFB or MAI

17-18

XIII. CSF

For CultureFor TB CultureFor Cryptococcus Antigen & Fungal Culture

19

XIV. Sputum

Collection of Sputum for CultureFor TB CultureFor Fungal Culture

20

XV. Corneal Scrapings 21

XVI. Tissue for Cultur e

For Anaerobic CultureFor TB CultureFor Fungal Culture

21


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I. Collection of Specimens for Microbio logy

Proper transport and storage of specimens are prerequisites for reliable cultureresults. Ensure that the requisition and specimen are labeled with:

patient's full name date of birth or health card number source of specimen

collection date and time

The examination requested should be specified on the requisition.

Information concerning anti-microbial therapy or allergy, pregnancy, clinical

diagnosis, or underlying disease should also be noted.

Specimens should be transported promptly to LifeLabs Laboratory. Delays toprocessing, beyond the recommended acceptable holding times, will compromise

culture results.

All specimens and body fluids are considered potentially hazardous. They shouldbe treated as if capable of causing disease. Clinical information or patient historyas provided by the physician determines the scope of the laboratory investigationby ensuring that all laboratory services are relevant or adequate for each clinicalsituation.


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I. II. Microbio logy Swab and Collection Containers

Sterile Cont ainer (100 mL) Urine Preservative/

Bor ic Acid Tube Pediatr ic Urin e

Collector

Swabs C&S- Amies Transpor t MediumCharcoal o r Clear Gel

PORT-A-CULanaerobic transport system

Aer obic Blue Anaerobic Go ld Pediat r ic Pi nk

Bactec Blood Culture Bottles Fungus Scraping Kit-For skin, hair or nails


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II. Microbio logy Swab and Collection Containers

Chlamy dia and GC Female- Pink Chlamyd ia and GC Male- Blu e

BD ProbeTec CT/GC Qx Ampli f ied DNA Assay Collection Kit

Stool Culture and Sensitiv ityCary B lair Medium

(pink l iquid)

Stool Ova and ParasitesSA F

(colour less l iquid)

Pinworm Col lect ion-Vials with Paddles


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III. Urine for Cultu re

Urines for culture should be collected as described below in a sterile 90 mLcontainer.

The patient's full name, date of birth or health card number, source of specimenand date and time of collection should be specified on the requisition and specimencontainer. Also include any information concerning pregnancy or antibiotic drugallergies on the requisition.

Collection of Midstream Urine Samples

Early morning urine specimens are preferred, although urine collected at othertimes of the day are acceptable. A mid-stream clean catch urine sample isnecessary for culture so that any bacteria present around the urethra and on thehands do not contaminate the specimen.

Use a sterile container for collection.

Complete the information requested on the container label- your full name, dateof birth or health card number, source of specimen and date and time ofcollection.

Wash and dry your hands thoroughly.

Remove the container lid and set it aside. Do not touch inner surfaces ofcontainer.

Wash your urogenital area (lower parts) with the towelette provided.

Women: wipe from front to back between the folds of skin.

Men: retract the foreskin (if un-circumcised), and clean the glans (head of the

penis). Pass a small amount of urine into the toilet (women: hold skin folds apart) and

then midway through urination, urinate into the container. The container shouldonly be 1/2 to 2/3 full.

Replace the lid and tighten firmly.

Wash and dry your hands thoroughly.

Immediately refr igerate the specimen and submit to the laboratory WITHIN24 hours of col lect ion (maintain at 2-8 C when transporting).

If transportation to the laboratory is expected to go beyond 24 hours, transfer10mL of urine into an NCS tube with boric acid preservative (#29323).

Maintain preserved urine (NCS tube) at room temperature and submit to thelaboratory WITHIN 72 hours of collection.

Pediatric urine collection kits are also available.

For TB Culture

Collect at least 40 mL catheter or midstream first morning urine specimens in asterile container for three consecutive days. Do not pool specimens.

Store refrigerated at 2-8C and submit each specimen to the PHL laboratory witha completed PHL requisition.


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IV. Swabs for Cultu re

Swabs from the genital tract, throat, eye, ear, nose, and superficial wounds (e.g.,sores, boils, and rashes) should be transported to the laboratory in the transport

medium provided by the local LifeLabs Laboratory (charcoal or clear gel transport

medium).

The patient's ful l name, date of birth or health card number, the anatomicalsite swabbed and date and time of col lection should be specified on therequisit ion and specimen container to assist in the evaluation of bacteriaisolated. Also include any information concerning pregnancy or antibiotic drugallergies or therapy on the requisition.

All swabs have expiry dates, so rotation of stock is important. Dry swabs received

for culture will not be processed unless received within one hour of collection.

Collection of Wound Swabs

Gentle cleansing of a skin wound prior to sample collection is recommended toreduce commensal flora contamination.

Purulent exudates must be expressed onto swabs.

Place the swab into the transport media.

Label swab with patients full name, date of birth or health card number,source of specimen and date and time of collection.

Maintain swabs at room temperature and submit to the laboratory WITHIN24 hours of col lect ion.

Deep wound specimens (aspirates) are optimal as the predictive value of

superficial swabs is low. Investigation of deep wounds for anaerobes requiresa special anaerobic collection kit, available from the LifeLabs laboratory.

Collectio n of Eye Swabs

Collect before topical or anesthetics are applied.

Swab pus or purulent discharge taken from the lower inverted lid.


Label swab with patients full name, date of birth or health card number,

source of specimen and date and time of collection.

Maintain swabs at room temperature and submit to the laboratory WITHIN

24 hours of col lect ion.

Collectio n of Ear Swabs Swab the external ear canal.




Maintain swabs refrigerated at 2-8 C and submit to the laboratory WITHIN



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IV. Swabs for Cultur e (Cont d)

Collection of Throat Swabs Swab the tonsillar area and/or posterior pharynx.



Maintain swabs at room temperature and submit to the laboratory WITHIN


Genital Swab Cultures inc lude

Organism/SyndromeInvestigated Source of Specimen Collection

Neisseria gonorrhoeae Endocervical, UrethralAmies Transport Swab

(Charcoal or clear gel)

Yeast, Bacterial Vaginosis,Trichomonas

Post Vaginal VaultAmies Transport Swab


Prenatal Screening forGroup B Streptococcus at35-37 weeks gestation

Combined Vaginal/RectalAmies Transport Swab


Collection of Urethral Specimens for Culture ofNeisseri a gono rrh oeae

Express exudates from the urethra and collect it on a swab.

If exudate is unavailable, insert an urethrogenital swab about 2 cm into the

urethra, gently rotate it and remove. Place the swab into the transport medium.



Maintain swabs at room temperature and submit to the laboratory WITHIN 24

hours of collection.

Collection of Endocervical Swabs for Culture ofNeisseri a gono rrh oeae

Moisten the speculum with warm water. Lubricants may be toxic to Neisseriagonorrhoeae. Remove any mucous or vaginal material.

Gently compress the cervix with the blades of the speculum. Collect the endocervical discharge onto the swab or insert the swab into the

cervix.

Place the swab into the transport medium.



Maintain swabs at room temperature and submit to the laboratory WITHIN 24hours of collection.


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IV. Swabs for Cultu re (Cont d)

Collection of Vaginal Swabs for Yeast, Bacterial Vaginosis & Trichomonas

Wipe away any excessive amount of secretion.

Obtain secretions from the mucosal membrane with a swab.

Place the swab into the transport medium.


Maintain swabs at room temperature and submit to the laboratory WITHIN 24

hours of collection.

Collectio n of Vaginal/Rectal Swabs for Prenatal Group B Streptoco ccus

Screening at 35-37 Weeks Gestation Do not use a speculum for collection.

Swab the lower vagina (vaginal introitus).

Using the same swab or a different swab, insert swab through the anal

sphincter to swab the rectum.

Place the swab(s) into the transport medium.



Maintain swabs at room temperature and submit to the laboratory WITHIN 24hours of collection.

Clearly indicate on the requisition that specimens are for Group B Streptococcalculture.

Requisition should indicate a penicillin allergy when applicable.


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V. Chlamydia trachomatis & Neisseria gonor rhoeae:

Nucleic Acid Amp lif ic ation Testing (NAAT)

Test Source of Specimen Collection

Chlamydia trachomatis &

Neisseria gonorrhoeae*

Nucleic Acid Ampli f icat ionTesting (NAAT)

Endocervical, Urethral,Vaginal

BD ProbeTec CT/GC QxAmplified DNA

Assay collection kit. (male blue;female pink).

Swab container contains liquid.

Urine 90 mL sterile container

*In patients < 14 years of age, specimens should be submitted for Neisseria gonorrhoeae culture

in charcoal or clear gel transport medium.

Specimens for Chlamydia trachomatis and Neisseria gonorrhoeae NAAT testing should

be collected, stored and submitted to the laboratory as described below.

The patient's ful l name, date of birth or health card number, the source of

specimen and date and time of col lection should be specified on the requisit ion

and specimen.

All swabs have expiry dates so rotation of stock is important.

NOTE:Testing fo r both Chlamydia trachomatis an d Neisseria go norrh oeae maybe perfor med on the same swab or urine specimen.

Collection of Endocervical Swab forChlamydia trachomatis &

Neisseria gono rrho eae NAAT

Use BD ProbeTec CT/GC Qx Amplified DNA Assay collection kit.(female pink).

Remove excess mucus and blood from the endocervix with the large tippedcleaning swab with the white shaft. Discard the cleaning swab.

Do not pre-wet the pink collection swab prior to collecting the specimen.

Insert the pink collection swab into cervical canal and rotate for 15-30 seconds.

Withdraw the swab carefully. Avoid contact with the vaginal mucosa.

Uncap the CT/GC Qx Swab Diluent tube.

Fully insert the pink collection swab into the CT/GC Q x Swab Diluent tube. Break

the shaft of the swab at the score mark. Use care to avoid splashing of contents. Ensure the cap is tightly secured.

Label tube with patients full name, date of birth or health card number, source of

specimen and date and time of collection.

Place each swab container into a separate Ziploc plastic baggie for transport to the

laboratory.

Store and ship at 2-8 C. Swabs are stable for 30 days.


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V. Chlamydia trachomatis & Neisseria gonor rhoeae: NAAT (Contd )

Collection of Urethral Swab for Chlamydia tr achomatis &

Neisseria gono rrho eae NAAT Use BD ProbeTec CT/GC Qx Amplified DNA Assay collection kit. (male- blue).

Do not pre-wet the blue collection swab prior to collecting the specimen.

Insert the blue collection swab 2-4 cm into the urethra and rotate for 3-5 seconds.

Withdraw the swab.

Uncap the CT/GC Qx Swab Diluent tube. Fully insert the blue collection swab into

the CT/GC Qx Swab Diluent tube. Break the shaft of the swab at the score mark.

Use care to avoid splashing of contents.

Ensure the cap is tightly secured.

Label tube with patients full name, date of birth or health card number, source ofspecimen and date and time of collection.

Place each swab container into a separate Ziploc plastic baggie for transport to the

laboratory.

Store and ship at 2-8 C. Swabs are stable for 30 days.

Collection of Urine for Chlamydia trachomatis & Neisseria gonorrhoeae NAAT

The patient should not have urinated for at least 1 hour prior to specimen collection.

Collect 20 to 60 mL of f irst catch urine (the first part of the stream- NOTmidstream) into a sterile 90 mL container.

Label the container with the patients full name, date of birth or health card number,


Indicate total volume of urine if 60mL is collected.

Store and ship 2-8C. Urines are stable for 7 days.

Note: Testing for both Chlamydia and Neisseria gonorrhoeae may be performed onthe same urine specimen.

A sep ar at e f i rs t cat ch ur ine sam ple is req ui red for molec ular tes t ing.If any other tests are required, e.g. C&S and urinalysis, then another specimen shouldbe collected for these tests.


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VI. Chlamydia Culture

Collection of Specimens for ChlamydiaCultureChlamydia Cultur e

Non-genital Swabs (rectal,eye, conjunctivial scrapingsand swabs, pharyngeal)

Chlamydia Culture Kit available fromPublic Health Laboratories

Specimens are collected by the physician in the Chlamydia Culture Kit provided byPublic Health Laboratory.

Label the container with the patients full name, date of birth or health card number,source of specimen and date and time of collection.

Specimen MUST be transported at 2-8 C on ice packs to PHL laboratory within 48hours of collection.

If longer storage is required, store and transport FROZEN at -70 C on DRY ICE.

VII. Surveil lance Specimens

Appropriate specimens for screening are:

MRSA: Nasal Swab, Rectal/ Perineal Swab, Axilla Swab, Wound Swab, Swabfrom sites previously positive for MRSA, Sputum, Urine, Stool

VRE: Rectal Swab, Perianal Swab, Swab from sites previously positive for VRE,Urine, Stool. Nasal swabs are not appropriate for VRE.

ESBL and CPE/CRE (Carbapenemase): Rectal Swab, Wound Swab, Swab fromsites previously positive, Urine, Stool, Endotracheal secretions or samples from

other exit sites Group A Streptococcus: Throat Swabs, Nasal Swabs, Rectal Swabs, Vaginal

Swabs and Skin Lesion Swabs

Record the target organism as well as body site on the requisition. E.g. Rectal for VRE.

Swabs should be collected and transported to the laboratory in the transport medium

provided by the local LifeLabs Laboratory (charcoal or clear gel transport medium).

For MRSA, VRE and Group A Streptococcus a separate swab should be collected for

each target organism requested.

Urines, stools, sputum, endotracheal secretions or samples from other exit sites

should be collected in sterile containers. Stools in Cary Blair container (white lid andpink fluid) container are also acceptable.

Refrigerate urine, stool and sputum specimens at 2-8C and submit to the laboratoryWITHIN 24 hours of collection.

Maintain all other specimens at room temperature and submit to laboratory WITHIN 24hours of collection.


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VIII. Stoo l Specimens

Collection of Stool Specimens

Avoid barium, oil or magnesium before a stool collection. Wash and dry your hands.

Urinate into the toilet if needed.

Lift the toilet seat. P lace sheets of plastic wrap (e.g., Saran wrap) over thetoilet bowl, leaving a slight dip in the center. P lace the toilet seat down.

Pass the stool onto the plastic wrap. OR Use a clean bowl or bedpan. Do notlet urine or water touch the stool specimen.

Open the container and transfer small portions of the stool into theappropriate container as described below. Do not overfill the container.

Replace the cap and firmly tighten.

Label the container with the patients full name, date of birth or health card

number, source of specimen and date and time of collection. Place in Ziploc bag and seal carefully.

Note that a leaking container may be INFECTIOUS.

Wash and dry hands thoroughly.

Indicate the date and time of collection on the requisition.

Stool Cul ture

To the Cary B laircontainer (white lid and pink fluid), add stool to the Fi l l L ine.Tighten cap and ensure that specimen is well mixed. Refrigerate specimen at 2-8C and submit to the laboratory WITHIN 24 hours of collection. Samples may beaccepted up to 48 hours after collection.

Multiple stool specimens collected on the same day from the same patient forC&S will not be processed. The first specimen received will be processed.

OAML Guideline (CLP-O19) recommends that physicians submit a singlestoo l specimen for C&S to determine the cause of acute diarrhea or to assessindividuals suspected of having been infected in an outbreak situation (foodpoisoning).

Ova and Parasites To the SAF fixative container (red lid and clear fluid), add 2-3 spoonfuls of stooluntil the liquid reaches the Fill Line. After replacing the cap, gently shake thecontainer until the specimen is well mixed with the liquid. Maintain specimen atroom temperature and submit to the laboratory WITHIN 72 hours of collection.

Caut ion: SAF FIXATIVE CONTAINS FORMALDEHYDEToxic by inhalat ion and i f swal lowed. I rr i tat ing to the eyes, respiratorysystem, and skin. May cause sensi t izat ion by inhalat ion or by s kin contact.Risk of serious d amage to eyes.

OAML guideline (CLP-O18) recommends that physicians initially request testingof only one properly collected stool specimen for ova and parasite examination.Subsequent testing should be requested only if clinically indicated and theresults of the first test are negative or if positive results do not match the clinicalsituation.


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VIII. Stool Specimens (Cont d)

Clostridium dif f ici le Toxin

Testing is performed at PHL. Send specimen with completed PHL requisition.Testing will not be performed on infants less than 12 months old.

To the sterile 90 mL container, add 2-3 teaspoonfuls (5 to 10 mL) of diarrhoealstool. Refrigerate specimen at 2-8C and submit to the laboratory WITHIN 72hours of collection.

For longer storage, freeze at -20 C. Keep specimen in freezer until ready forpickup. Ship to PHL on a frozen ice pack. Record the date frozen on PHLrequisition.

IX. Parasitolo gy Specimens

Collection for Pinworm Examination

A special kit that includes detailed collection instructions for the patient isavailable from the local LifeLabs Laboratory.

The specimen must be collected early in the morning before bathing or usingthe toilet. The sticky surface of the specimen paddle is pressed firmly againstthe skin surrounding the anus to pick up eggs.

Return the paddle to the transport container provided.

Label container with patient's full name, date of birth or health card number,and collection date and time.

Maintain refrigerated at 2-8C and submit to the laboratory WITHIN 48 hours ofcollection.

Multiple consecutive samples are usually required to diagnose infection as

deposition of the eggs on the perianal skin is sporadic.

Collection of Urine for Schistosom a

Collect the last voided portion of mid-day urine sample into a sterile 90 mLcontainer.

Label the container with patients full name, date of birth or health cardnumber, source of specimen and date and time of collection.

Maintain at room temperature and submit to the laboratory WITHIN 24 hours ofcollection.

Collection of Specimens f or Identif i cation (Non-Stool): Worm, Insect,Proglottid, Skin Scrapings for Scabies, Hair Specimens f or L ice

Testing is performed at PHL. Send specimen with completed PHL requisition.

Specimens include worms, insects, proglottids, skin scrapings for scabies, hair

specimens for lice and unknown substances.

Place into a sterile 90 mL container.

Label the container with patients full name, date of birth or health cardnumber, source of specimen and date and time of collection.



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X. Fungus/Mycology for Dermatophytes

Skin scraping, hair, and nail clipping should be submitted wrapped in heavy

black paper. Collection kits are available from the local LifeLabs Laboratory.

Vaginal, throat, mouth and ear swabs are collected in Amies' transport media.

The patient's full name, date of birth or health card number, specimen source

and date and time of collection should be specified on the requisition and

specimen container.

Fungal cultures for dermatophytes require up to four weeks of incubation.

Nail, Skin and Hair Specimens

Store at room temperature and submit to the laboratory WITHIN 72 hours ofcollection.

Vaginal, Throat, Mout h and Ear Swabs

Store ear swabs at 2-8C and all other swabs at room temperature. Submit to the

laboratory WITHIN 24 hours of collection.

Collection of Nail Specimen

Cleanse the nail area with 70% alcohol to eliminate contaminating body

bacteria. Allow to air dry.

The part of the nail that is the optimal specimen will vary with the type of

onychomycosis. For distal subungual infection, the healthy nail plate should be

discarded. A sample from the nail bed and from material from underside the

nail plate should be obtained. For proximal subungual onychomycosis, the

healthy nail plate should be removed and material from the proximal nail bed

obtained. For white superficial onychomycosis, material from the white area of

the nail plate should be obtained.

A scalpel blade or a small curet can be used to obtain the sample. Submit in a

sterile container.


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XI. Sterile Fluids (Other than Blood and CSF)

Aseptic collection technique is required in the sampling of sterile body sites.Fluids include: peritoneal, pleural and pericardial, joint, bile, and aspirates.

The patient's full name, date of birth or health card number, the source of thefluid and date and time of col lection should be specified on the requisition andspecimen container.

Fluids for culture should be submitted as described below. Provide any relevantclinical information on the requisition.

In order to avoid needle st ick in jury in the laboratory, specimens includingneedles wil l not be accepted.

Request Collection Container Instruc tion s

C& S

MinimumQuantity

1 mL

Routine:Sterile 90 mL container or red top tube.

Collection of fluids into tubes with anticoagulant is notrecommended but will be processed if received in a vacutainerwith anticoagulant. Fluid collected in a tube with heparin (greentop) is preferred over citrate (blue top) or EDTA (lavender top)

If anaerobic workup is specifically requested: AnaerobicTransport system + screw top container (for Gram)

Synovial, peritoneal, joint, ascites, pleural and pericardial fluidsare acceptable for culture in BACTEC blood culture bottles. Ifsterile body fluids are received in BACTEC bottles, they should

be accompanied by a specimen in a sterile container for Gramstain.

Synovial/J oint Fluid:If < 2 mL, use Pediatric Blood Culture Bottle.If > 2 mL, use Aerobic Blood Culture Bottle. If anaerobic infectionsuspected, add an additional Anaerobic Blood Culture Bottle.

Peritoneal Fluid, Pericardial Fluid, Pleural Fluid, Ascites Fluid:

Inoculate a minimum of 3 mL into an Aerobic and Anaerobic

Blood Culture Bottle.

Store and ship atRoom Temperature

Exception: PericardialFluids must berefrigerated at 2-8C

Ideally specimenshould be submittedwithin 12 hours ofcollection.

However within 24

hours of collection isacceptable.

TB

MinimumQuantity

1 mL

Collect as much as possible (10-15 mL) in a sterile 90 mLcontainer.Submit with completed PHL requisition.

Store at RoomTemperature

Submit to the

laboratory WITHIN 24hours

Fungus

(not CSF)

MinimumQuantity

1 mL

Sterile 90 mL container.Submit with completed PHL requisition.

Store at RoomTemperature.

Submit to thelaboratory WITHIN 24hours of collection


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XII. Blood Cultu res

LifeLabs provides special blood culture bottles upon request. Patient blood isdrawn directly into these containers, mixed well and sent directly to thelaboratory.

A sufficient volume of blood withdrawn per culture is one of the most importantfeatures to optimize recovery of organisms from blood. Mark each bottle labelbefore collection with the desired fill line to indicate the desired collectionvolume. Overfilling of bottles should be avoided.

The patient's full name, date of birth or health card number and date and timeof col lect ion should be specified on the requisition an d blood culture bottle.Also include on the requisition any antib iot ics in use and important cl in icalh is tory (i.e. fever of unknown origin, query endocarditis).

Maintain Blood cultures at room temperature. Optimally submit tolaboratory WITHIN 12 hours of col l ection.

Specimens are adequate if submitted within 24 hours of collection.

Adul t Co l lec t ion

Requests Collection Procedure

Routine Blood

Cultures

Collect two (2) sets of blood cultures.

1st

set: 10 mL of blood collected into 1 aerobic (blue) an d 10 mL ofblood collected into 1 anaerobic (gold) BACTEC bottle.

2nd set: 10 mL of blood collected into 1 aerobic (blue) BACTECbottle collected from a 2

ndvenipuncture site.

There is no need to wai t between col lect ion of sets of bloodcultures unless endocarditis is suspected (see below).

Endocardi t is,

SBE, Bartonella,

Bruc ella** or

HACEK

**Notif y medical

microbiologist i f Brucella requested.

Collect three sets of blood cultures.

Each set should be at least 30 minutes apart.

1st

set: 10 mL of blood collected into 1 aerobic (blue) and 10 mL ofblood collected into 1 anaerobic (gold) BACTEC bottle.

2nd

set: 10 mL of blood collected into 1 aerobic (blue) and 10 mL ofblood collected into 1 anaerobic (gold) BACTEC bottle from

a 2nd venipuncture site.

3rd

set: 10 mL of blood collected into 1 aerobic (blue) and 10 mL ofblood collected into 1 anaerobic (gold) BACTEC bottle froma 3

rdvenipuncture site


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XII. Blood Cultures (contd)

Child Collection

WeightApproximat e

Age

Total Volumeof Blood

Collection Procedure

< 2 kg(< 5 lb)

Neonate(< 1 month)

1 to 2 mL 1 BACTEC PEDS (pink)

2.1 - 12.7 kg

(5 to 28 lb)

Infant(1 month to 2 years)

3 to 5 mL 1 BACTEC PEDS (pink)

12.8 - 36.3 kg

(28 to 80 lb)

Children

(2 to 12 years)

5 to 10 mL

(5 mL per bottle)

2 BACTEC PEDS (pink)bottles collected from samevenipuncture site

> 36.3 kg

(> 80 lb)

Adolescent

(> 12 years)

20 mL

(10 mL per bottle)

1 aerobic BACTEC bottle (blue)1 anaerobic BACTEC bottle(gold)

bottles collected from samevenipuncture site

Request f or Fungu s, Yeast, Cryptococcus, Mycobacteria, TB, AFB or MAI

Requests Collection Procedure

Fungi/Yeast

Cryptococcus

Candida

Two sets:

1st set = 10 mL of blood collected into 1 aerobic (blue) and 10 mL ofblood collected into 1 anaerobic (gold) BACTEC bottle.

2nd set = 10 mL of blood collected into 1 aerobic (blue) BACTEC bottleand 10 mL of blood collected into a 2nd aerobic (blue)BACTEC bottle collected from a second venipuncture site.

Dimorphic Fungi:

Histoplasmosis

Blastomycosis

Coccidiodes

Mould: Aspergi llus

Zygomycetes

Malassezia furfur

Sodium Heparin (green stopper) vacutainer tube (7 mL)

Maintain at room temperature. Submit to PHL within 24 hours with acompleted PHL requisition.

Mycobacteria /TB/

AFB/MAI


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XIII. CSF

For C&S Collect the CSF specimen into sterile container or red top tubes.

A minimum volume of 0.5 mL is recommended for routine culture.

Ensure that the specimen and requisit ion are labeled with the patient'sful l name, date of birth or health card number, source of specimen anddate and time of col lectio n.

CSF specimens should be maintained at room temperature. Do notrefrigerate.

Specimens should be both transported to the laboratory and processed asexpediently as possible.

Physicians submitt ing CSF specimens outside of a hospi tal sett ing

should not i fy the local L i feLabs Laboratory in advance.

For TB Culture

Collect specimen into sterile container or red top tube.

Volume should be at least 2 mL, samples less than 0.5 mL will be rejected.

Maintain at room temperature. Submit to PHL within 24 hours with acompleted PHL requisition.

For Cryptococcus A ntigen and Fungal Culture

Collect specimen into sterile container or red top tube.

Send a minimum of 0.5 mL CSF.

Complete PHL requisition requesting STAT testing for Cryptococcal antigenand fungus culture.

Store refrigerated at 2-8C and submit to the PHL laboratory as soon aspossible.


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XIV. Sputum

Early morning sputum samples are preferable for routine C&S, but samples

collected at other times of the day are also acceptable. Sputum is materialbrought up from as far down in the lungs as possible. It is not saliva.

Collect the sample into a 90 mL sterile container.

Ensure that the specimen and requisition are labeled with the patient's fullname, date of birth or health card number, source of specimen and date andtime of collection.

***Physicians requiring investigation of respiratory specimens from cysticfibrosis patients are advised to send patients to CF facilities for specimencollection and processing.

Collection of Sputum Samples for Culture

Wash and dry your hands.

If you wear dentures, remove them.

Rinse your mouth with water.

Remove the container cap.

After a big cough, place the sputum into the empty 90 mL sterile container.

Carefully and tightly replace the cap.

Wash your hands after collecting the specimen.

Store refrigerated at 2-8C and submit to the laboratory WITHIN 24 hours ofcollection.

Special requests for TB, fungus/mycology or other requests should be noted onthe requisition. Samples collected early in the morning are optimal.

A separate sputum specimen is optimal for each request (i.e., one sample forroutine bacteriology, one for TB and one for mycology, if all 3 requests areindicated).

For TB Culture

Do not rinse mouth with water as saprophytic mycobacteria in tap water mayproduce false positive results.

Collect an early morning specimen from a deep, productive cough on threeconsecutive days. Do not pool specimens.

Store refrigerated at 2-8C and submit to the PHL laboratory WITHIN 24hours of collection with a completed PHL requisition.

For Fungal Culture

Collect an early morning specimen.

Store refrigerated at 2-8C and submit to the PHL laboratory WITHIN 24hours of collection with a completed PHL requisition.


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XV. Corneal Scrapings

Corneal scrapings are obtained by the physician using aseptic technique andideally should be inoculated at the time of collection directly onto theappropriate media and slides for the diagnosis of keratitis.

Ophthalmologists should contact LifeLabs at 416-675-3637 or 877-849-3637 atleast 24 hours in advance for approval and shipment of appropriate media andslides.

XVI. Tissue for Cultur e

Tissue for routine C&S should be collected in a sterile container. For smallsamples add several drops of sterile saline to the container to maintainmoisture.


A separate sample is optimal for each request

For Anaerobic Culture

If anaerobic culture is required then submit the tissue in a PORT-A-CULanaerobic transport system.

Store at room temperature and submit to the laboratory WITHIN 24 hours ofcollection.

For TB Culture

Collect in a sterile container.

Store at room temperature and submit to the PHL laboratory WITHIN 24 hours ofcollection with a completed PHL requisition.

For Fungal Culture

Collect in a sterile container.Store at room temperature and submit to the PHL laboratory WITHIN 24 hours ofcollection with a completed PHL requisition.

06 microbiology - specimen handling collection instructions

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