MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS

TABLE OF CONTENTS I. II. III. Collection of Specimens for Microbiology Microbiology Swab and Collection Containers Urine for Culture
Collection of Midstream Urine Samples Urine for TB Culture

Page 3 4-5 6

IV.

Swabs for 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 of Neisseria gonorrhoeae Collection of Vaginal Swabs Collection of Vaginal/Rectal Swabs for Prenatal Group B Streptococcus Screening at 35-37 Weeks Gestation

7-9

V.

Chlamydia trachomatis : Nucleic Acid Amplification Testing & Neisseria gonorrhoeae : Nucleic Acid Amplification Testing
Collection of Endocervical Swab for CT/ N. gonorrhoeae NAAT Collection of Urethral Swab for CT/ N. gonorrhoeae NAAT Collection of Urine for CT/ N. gonorrhoeae NAAT

10-11

VI.

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

12

VII. VIII.

Surveillance Specimens Stool Specimens

Collection of Stool Specimens Stool Culture Ova and Parasites Clostridium difficile Toxin

12 13-14

IX.

Parasitology Specimens
Collection for Pinworm Examination Collection of Urine for Schistosoma Collection of Specimens for Identification (non-stool): W orms, Insect, Proglottid, Skin Scrapings for Scabies, Hair Specimens for Lice

14

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS

TABLE OF CONTENTS X. Fungus/Mycology for Dermatophytes
Nail, Skin and Hair Specimens Vaginal, Throat, Mouth and Ear Swabs Collection of Nail Specimen

Page 15

XI. XII.

Sterile Fluids (Other than Blood & CSF) Blood Cultures

Adult Collection Child Collection Request for Fungus, Yeast, Cryptococcus, Mycobacteria, TB, AFB or MAI

16 17-18

XIII.

CSF
For Culture For TB Culture For Cryptococcus Antigen & Fungal Culture

19

XIV.

Sputum
Collection of Sputum for Culture For TB Culture For Fungal Culture

20

XV. XVI.

Corneal Scrapings Tissue for Culture

For Anaerobic Culture For TB Culture For Fungal Culture

21 21

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS I. Collection of Specimens for Microbiology
Proper transport and storage of specimens are prerequisites for reliable culture results. 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 to processing, beyond the recommended acceptable holding times, will compromise culture results. All specimens and body fluids are considered potentially hazardous. They should be treated as if capable of causing disease. Clinical information or patient history as provided by the physician determines the scope of the laboratory investigation by ensuring that all laboratory services are relevant or adequate for each clinical situation.

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS II. Microbiology Swab and Collection Containers

I.

Sterile Container (100 mL)

Urine Preservative/ Boric Acid Tube

Pediatric Urine Collector

Swabs C&S- Amies Transport Medium Charcoal or Clear Gel

PORT-A-CUL anaerobic transport system

Aerobic Blue

Anaerobic Gold

Pediatric Pink

Bactec Blood Culture Bottles

Fungus Scraping KitFor skin, hair or nails

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS

II.

Microbiology Swab and Collection Containers

Chlamydia and GC Female- Pink

Chlamydia and GC Male- Blue

BD ProbeTec CT/GC Q x Amplified DNA Assay Collection Kit

Stool Culture and Sensitivity Cary Blair Medium (pink liquid)

Stool Ova and Parasites SAF (colourless liquid)

Pinworm CollectionVials with Paddles

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS III. Urine for Culture
Urines for culture should be collected as described below in a sterile 90 mL container. The patient's full name, date of birth or health card number, source of specimen and date and time of collection should be specified on the requisition and specimen container. Also include any information concerning pregnancy or antibiotic drug allergies on the requisition.

Collection of Midstream Urine Samples

Early morning urine specimens are preferred, although urine collected at other times of the day are acceptable. A mid-stream clean catch urine sample is necessary for culture so that any bacteria present around the urethra and on the hands do not contaminate the specimen. Use a sterile container for collection. Complete the information requested on the container label- your full name, date of birth or health card number, source of specimen and date and time of collection. Wash and dry your hands thoroughly. Remove the container lid and set it aside. Do not touch inner surfaces of container. 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 should only be 1/2 to 2/3 full. Replace the lid and tighten firmly. Wash and dry your hands thoroughly. Immediately refrigerate the specimen and submit to the laboratory WITHIN 24 hours of collection (maintain at 2-8 C when transporting). If transportation to the laboratory is expected to go beyond 24 hours, transfer 10 mL of urine into an NCS tube with boric acid preservative (#29323). Maintain preserved urine (NCS tube) at room temperature and submit to the laboratory 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 a sterile container for three consecutive days. Do not pool specimens. Store refrigerated at 2-8C and submit each specimen to the PHL laboratory with a completed PHL requisition.
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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS IV. Swabs for Culture
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 full name, date of birth site swabbed and date and time of requisition and specimen container isolated. Also include any information allergies or therapy on the requisition. or health card number, the anatomical collection should be specified on the to assist in the evaluation of bacteria concerning pregnancy or antibiotic drug

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 to reduce 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 WITHIN 24 hours of collection. Deep wound specimens (aspirates) are optimal as the predictive value of superficial swabs is low. Investigation of deep wounds for anaerobes requires a special anaerobic collection kit, available from the LifeLabs laboratory. Collect before topical or anesthetics are applied. Swab pus or purulent discharge taken from the lower inverted lid. 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 WITHIN 24 hours of collection. Swab the external ear canal. 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 refrigerated at 2-8 C and submit to the laboratory WITHIN 24 hours of collection.

Collection of Eye Swabs

Collection of Ear Swabs

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS IV. Swabs for Culture (Contd)
Collection of Throat Swabs

Swab the tonsillar area and/or posterior pharynx. 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 WITHIN 24 hours of collection.

Genital Swab Cultures include

Organism/Syndrome Investigated
Neisseria gonorrhoeae Yeast, Bacterial Vaginosis, Trichomonas Prenatal Screening for Group B Streptococcus at 35-37 weeks gestation

Source of Specimen
Endocervical, Urethral Post Vaginal Vault Combined Vaginal/Rectal

Collection
Amies Transport Swab (Charcoal or clear gel) Amies Transport Swab (Charcoal or clear gel) Amies Transport Swab (Charcoal or clear gel)

Collection of Urethral Specimens for Culture of Neisseria gonorrhoeae

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. 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 collection.

Collection of Endocervical Swabs for Culture of Neisseria gonorrhoeae

Moisten the speculum with warm water. Lubricants may be toxic to Neisseria gonorrhoeae . 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. 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 collection.
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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS IV. Swabs for Culture (Contd)
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. 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 collection.

Collection of Vaginal/Rectal Swabs for Prenatal Group B Streptococcus 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. 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 collection. Clearly indicate on the requisition that specimens are for Group B Streptococcal culture. Requisition should indicate a penicillin allergy when applicable.

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS V. Chlamydia trachomatis & Neisseria gonorrhoeae : Nucleic Acid Amplification Testing (NAAT)
Test Source of Specimen
Endocervical, Urethral, Vaginal

Collection
BD ProbeTec CT/GC Q x Amplified DNA Assay collection kit. (male blue; female pink). Swab container contains liquid.

Chlamydia trachomatis & Neisseria gonorrhoeae * Nucleic Acid Amplification Testing (NAAT)

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 full name, date of birth or health card number, the source of specimen and date and time of collection should be specified on the requisition and specimen. All swabs have expiry dates so rotation of stock is important. NOTE: Testing for both Chlamydia trachomatis and Neisseria gonorrhoeae may be performed on the same swab or urine specimen.

Collection of Endocervical Swab for Chlamydia trachomatis & Neisseria gonorrhoeae NAAT

Use BD ProbeTec CT/GC Q x Amplified DNA Assay collection kit. ( female pink ). Remove excess mucus and blood from the endocervix with the large tipped cleaning 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 Q x 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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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS

V. Chlamydia trachomatis & Neisseria gonorrhoeae : NAAT (Contd)

Collection of Urethral Swab for Chlamydia trachomatis & Neisseria gonorrhoeae NAAT

Use BD ProbeTec CT/GC Q x 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 Q x Swab Diluent tube. Fully insert the blue 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.

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 first catch urine (the first part of the stream- NOT midstream) into a sterile 90 mL container. Label the container with the patients full name, date of birth or health card number, source of specimen and date and time of collection. Indicate total volume of urine if <20mL or >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 on
the same urine specimen. A separate first catch urine sample is required for molecular testing. If any other tests are required, e.g. C&S and urinalysis, then another specimen should be collected for these tests.

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS VI. Chlamydia Culture

Collection of Specimens for Chlamydia Culture
Chlamydia Culture Non-genital Swabs (rectal, Chlamydia Culture Kit available from eye, conjunctivial scrapings Public Health Laboratories and swabs, pharyngeal)

Specimens are collected by the physician in the Chlamydia Culture Kit provided by Public 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 48 hours of collection. If longer storage is required, store and transport FROZEN at -70 C on DRY ICE.

VII. Surveillance Specimens

Appropriate specimens for screening are: MRSA: Nasal Swab, Rectal/ Perineal Swab, Axilla Swab, Wound Swab, Swab from 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 from sites 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 and pink fluid) container are also acceptable. Refrigerate urine, stool and sputum specimens at 2-8C and submit to the laboratory WITHIN 24 hours of collection. Maintain all other specimens at room temperature and submit to laboratory WITHIN 24 hours of collection.

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS VIII. Stool 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. Place sheets of plastic wrap (e.g., Saran wrap ) over the toilet bowl, leaving a slight dip in the center. Place the toilet seat down. Pass the stool onto the plastic wrap. OR Use a clean bowl or bedpan. Do not let urine or water touch the stool specimen. Open the container and transfer small portions of the stool into the appropriate 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 Culture To the Cary Blair container (white lid and pink fluid), add stool to the Fill Line . Tighten cap and ensure that specimen is well mixed. Refrigerate specimen at 28C and submit to the laboratory WITHIN 24 hours of collection. Samples may be accepted up to 48 hours after collection. Multiple stool specimens collected on the same day from the same patient for C&S will not be processed. The first specimen received will be processed. OAML Guideline (CLP-O19) recommends that physicians submit a single stool specimen for C&S to determine the cause of acute diarrhea or to assess individuals suspected of having been infected in an outbreak situation (food poisoning). Ova and Parasites To the SAF fixative container (red lid and clear fluid), add 2-3 spoonfuls of stool until the liquid reaches the Fill Line. After replacing the cap, gently shake the container until the specimen is well mixed with the liquid. Maintain specimen at room temperature and submit to the laboratory WITHIN 72 hours of collection.
Caution: SAF FIXATIVE CONTAINS FORMALDEHYDE Toxic by inhalation and if swallowed. Irritating to the eyes, respiratory system, and skin. May cause sensitization by inhalation or by skin contact. Risk of serious damage to eyes.

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

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS VIII. Stool Specimens (Contd)
Clostridium difficile 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 diarrhoeal stool. Refrigerate specimen at 2-8C and submit to the laboratory WITHIN 72 hours of collection. For longer storage, freeze at -20 C. Keep specimen in freezer until ready for pickup. Ship to PHL on a frozen ice pack. Record the date frozen on PHL requisition.

IX.

Parasitology Specimens
Collection for Pinworm Examination

A special kit that includes detailed collection instructions for the patient is available from the local LifeLabs Laboratory. The specimen must be collected early in the morning before bathing or using the toilet. The sticky surface of the specimen paddle is pressed firmly against the 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 of collection. Multiple consecutive samples are usually required to diagnose infection as deposition of the eggs on the perianal skin is sporadic.

Collection of Urine for Schistosoma

Collect the last voided portion of mid-day urine sample into a sterile 90 mL container. Label the container with patients full name, date of birth or health card number, source of specimen and date and time of collection. Maintain at room temperature and submit to the laboratory WITHIN 24 hours of collection.

Collection of Specimens for Identification (Non-Stool): Worm, Insect, Proglottid, Skin Scrapings for Scabies, Hair Specimens for Lice

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 card number, source of specimen and date and time of collection. Maintain at room temperature and submit to the laboratory WITHIN 24 hours of collection.
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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS 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 of collection. Vaginal, Throat, Mouth 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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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS

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 the fluid and date and time of collection should be specified on the requisition and specimen container. Fluids for culture should be submitted as described below. Provide any relevant clinical information on the requisition. In order to avoid needle stick injury in the laboratory, specimens including needles will not be accepted. Request
C&S Minimum Quantity 1 mL

Collection Container
Routine: Sterile 90 mL container or red top tube. Collection of fluids into tubes with anticoagulant is not recommended but will be processed if received in a vacutainer with anticoagulant. Fluid collected in a tube with heparin (green top) is preferred over citrate (blue top) or EDTA (lavender top) If anaerobic workup is specifically requested: Anaerobic Transport system + screw top container (for Gram) Synovial, peritoneal, joint, ascites, pleural and pericardial fluids are acceptable for culture in BACTEC blood culture bottles. If sterile body fluids are received in BACTEC bottles, they should be accompanied by a specimen in a sterile container for Gram stain. Synovial/Joint Fluid: If < 2 mL, use Pediatric Blood Culture Bottle. If > 2 mL, use Aerobic Blood Culture Bottle. If anaerobic infection suspected, 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.

Instructions
Store and ship at Room Temperature Exception: Pericardial Fluids must be refrigerated at 2-8C Ideally specimen should be submitted within 12 hours of collection. However within 24 hours of collection is acceptable.

TB Minimum Quantity 1 mL Fungus (not CSF) Minimum Quantity 1 mL

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

Store at Room Temperature Submit to the laboratory WITHIN 24 hours Store at Room Temperature. Submit to the laboratory WITHIN 24 hours of collection

Sterile 90 mL container. Submit with completed PHL requisition.

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS XII. Blood Cultures

LifeLabs provides special blood culture bottles upon request. Patient blood is drawn directly into these containers, mixed well and sent directly to the laboratory. A sufficient volume of blood withdrawn per culture is one of the most important features to optimize recovery of organisms from blood. Mark each bottle label before collection with the desired fill line to indicate the desired collection volume. Overfilling of bottles should be avoided. The patient's full name, date of birth or health card number and date and time of collection should be specified on the requisition and blood culture bottle. Also include on the requisition any antibiotics in use and important clinical history (i.e. fever of unknown origin, query endocarditis). Maintain Blood cultures at room temperature. laboratory WITHIN 12 hours of collection. Optimally submit to

Specimens are adequate if submitted within 24 hours of collection.

Adult Collection Requests

Routine Blood Cultures

Collection Procedure
Collect two (2) sets of blood cultures. 1 st set: 10 mL of blood collected into 1 aerobic (blue) and 10 mL of blood collected into 1 anaerobic (gold) BACTEC bottle. 2 nd set: 10 mL of blood collected into 1 aerobic (blue) BACTEC bottle collected from a 2 nd venipuncture site. There is no need to wait between collection of sets of blood cultures unless endocarditis is suspected (see below).

Endocarditis, SBE, Bartonella, Brucella** or HACEK **Notify medical microbiologist if Brucella requested.

Collect three sets of blood cultures. Each set should be at least 30 minutes apart. 1 st set: 10 mL of blood collected into 1 aerobic (blue) and 10 mL of blood collected into 1 anaerobic (gold) BACTEC bottle. 2 nd set: 10 mL of blood collected into 1 aerobic (blue) and 10 mL of blood collected into 1 anaerobic (gold) BACTEC bottle from a 2 nd venipuncture site. 3 rd set: 10 mL of blood collected into 1 aerobic (blue) and 10 mL of blood collected into 1 anaerobic (gold) BACTEC bottle from a 3 rd venipuncture site

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS XII. Blood Cultures (contd)
Child Collection
Weight
< 2 kg (< 5 lb) 2.1 - 12.7 kg (5 to 28 lb) 12.8 - 36.3 kg (28 to 80 lb)

Approximate Age
Neonate (< 1 month) Infant (1 month to 2 years) Children (2 to 12 years)

Total Volume of Blood

1 to 2 mL

Collection Procedure

1 BACTEC PEDS (pink)

3 to 5 mL

1 BACTEC PEDS (pink)

5 to 10 mL (5 mL per bottle)

2 BACTEC PEDS (pink) bottles collected from same venipuncture site 1 aerobic BACTEC bottle (blue) 1 anaerobic BACTEC bottle(gold) bottles collected from same venipuncture site

> 36.3 kg (> 80 lb)

Adolescent (> 12 years)

20 mL (10 mL per bottle)

Request for Fungus, Yeast, Cryptococcus, Mycobacteria, TB, AFB or MAI

Requests
Fungi/Yeast
Cryptococcus Candida

Collection Procedure
Two sets: 1st set = 10 mL of blood collected into 1 aerobic (blue) and 10 mL of blood collected into 1 anaerobic (gold) BACTEC bottle. 2nd set = 10 mL of blood collected into 1 aerobic (blue) BACTEC bottle and 10 mL of blood collected into a 2nd aerobic (blue) BACTEC bottle collected from a second venipuncture site.

Dimorphic Fungi: Histoplasmosis Blastomycosis Coccidiodes Mould: Aspergillus Zygomycetes Malassezia furfur

Sodium Heparin (green stopper) vacutainer tube (7 mL) Maintain at room temperature. Submit to PHL within 24 hours with a completed PHL requisition.

Mycobacteria /TB/ AFB/MAI

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS 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 requisition are labeled with the patient's full name, date of birth or health card number, source of specimen and date and time of collection. CSF specimens should be maintained at room temperature . Do not refrigerate. Specimens should be both transported to the laboratory and processed as expediently as possible. Physicians submitting CSF specimens outside of a hospital setting should notify the local LifeLabs 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 a completed PHL requisition.

For Cryptococcus Antigen 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 antigen and fungus culture. Store refrigerated at 2-8C and submit to the PHL laboratory as soon as possible.

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS 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 material brought 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 full name, date of birth or health card number, source of specimen and date and time of collection. ***Physicians requiring investigation of respiratory specimens from cystic fibrosis patients are advised to send patients to CF facilities for specimen collection 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 of collection. Special requests for TB, fungus/mycology or other requests should be noted on the requisition. Samples collected early in the morning are optimal. A separate sputum specimen is optimal for each request (i.e., one sample for routine bacteriology, one for TB and one for mycology, if all 3 requests are indicated).

For TB Culture
Do not rinse mouth with water as saprophytic mycobacteria in tap water may produce false positive results. Collect an early morning specimen from a deep, productive cough on three consecutive days. Do not pool specimens. Store refrigerated at 2-8C and submit to the PHL laboratory WITHIN 24 hours of collection with a completed PHL requisition. Collect an early morning specimen. Store refrigerated at 2-8C and submit to the PHL laboratory WITHIN 24 hours of collection with a completed PHL requisition.

For Fungal Culture

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MICROBIOLOGY SPECIMEN HANDLING & COLLECTION INSTRUCTIONS XV. Corneal Scrapings

Corneal scrapings are obtained by the physician using aseptic technique and ideally should be inoculated at the time of collection directly onto the appropriate media and slides for the diagnosis of keratitis. Ophthalmologists should contact LifeLabs at 416-675-3637 or 877-849-3637 at least 24 hours in advance for approval and shipment of appropriate media and slides.

XVI.

Tissue for Culture

Tissue for routine C&S should be collected in a sterile container. For small samples add several drops of sterile saline to the container to maintain moisture. Maintain at room temperature and submit to the laboratory WITHIN 24 hours of collection. A separate sample is optimal for each request

For Anaerobic Culture

If anaerobic culture is required then submit the tissue in a PORT-A-CUL anaerobic transport system. Store at room temperature and submit to the laboratory WITHIN 24 hours of collection.

For TB Culture
Collect in a sterile container. Store at room temperature and submit to the PHL laboratory WITHIN 24 hours of collection 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 of collection with a completed PHL requisition.

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