Virology

To expedite specimens to the lab and avoid delays in processing, it is essential to provide complete information with each specimen:

• Patient Name & Age
• Date of Collection
• Specimen Source
• Clinical Symptoms or Suspected Diagnosis
• Test(s) Requested by Numerical Order Code and Test Name

Pack only one specimen per specimen bag for transport.

General notes:

1. Use the correct specimen collection kit for the test methodology requested. Do not use wooden swabs, that may inhibit chlamydia. Do not use calcium alginate swabs, which may inhibit chlamydia and herpes virus. After collection, break swab tips off into tubes of media.
2. Collect the specimen aseptically.
3. Collect the specimen from the appropriate site within several days of onset of clinical symptoms.
4. Unless otherwise noted, viral and chlamydia culture specimens are stored and preferably transported refrigerated up to 3 days from date of collection.
5. Viruses vary in stability and some cannot be frozen. These viruses can be frozen only if the specimen is transported in VCM or equivalent: Varicella / Herpes zoster, RSV, measles, CMV. Do not freeze specimens for these viruses if collected in any transport media besides VCM or equivalent. Stability is limited to 3 days maximum.
6. Specimen Rejection Criteria:

• Specimens not in MCV or equivalent, stored/transported frozen for Varicella /Herpes zoster, RSV, measles, CMV
• Specimens in viral transport medium with penicillin for Chlamydia culture
• Diapers
• Dry swabs
• Gel-based Swabs
• Specimens with insufficient volume to perform test requested
• Blood collected in isolator tubes or any type of cell lysing system, BacTec or bacterial blood culture bottles
• Penile or semen specimen for HPV analysis
• Specimens not collected or transported according to instruction or specification
• Leaking specimens or specimens sent with needles

VCM or equivalent = Viral-Chlamydial-Mycoplasma transport medium (green-cap) available from client supplies

Chlamydia culture

Swab (cervix, urethral, rectal, conjunctiva, vaginal):  Do NOT use wooden or cotton swabs. Use sterile swab to remove mucous or exudate and discard. Use fresh sterile dacron swabs to obtain columnar/ cuboidal epithelial cells. Use VCM or equivalent. May freeze at -70°C (dry ice) in VCM or equivalent up to 5 days.

Urine:   Culture methodology is not recommended for this specimen type. Use nucleic acid amplification methodology such as PCR or TMA.Chlamydia DFA (Direct Fluorescent Antibody)

Test Slide:   Use specimen collection kit for Chlamydia trachomatis Direct Test (Slide). Store at room temperature or refrigerate. Stable up to a week.

Chlamydia probe (GenProbe)

Swab only: Use GenProbe Specimen Collection Kit for Urethral/Conjunctival specimens or GenProbe Specimen Collection Kit for Cervical specimens. Only female endocervical, male urethral or conjunctival swabs are acceptable. Store at room temperature or refrigerated. Stable up to a week. Do not freeze.

Viral DFA (Direct Fluorescent Antibody, slide test)

Slide: Obtain cellular material with sterile swab and transfer to clean glass slide in a circle smaller than 15-mm diameter or use cytospin preparation. (Provide 2 slides with one circle each or I slide with two circles for Influenza A& B. Provide 3 slides with one circle each or 1 slide with 3 circles for Parainfluenza 1, 2, 3). Store at room temperature. Stable up to one week.

Swab: Collect cellular material aseptically. May use VCM or equivalent. Store refrigerated.

Gastrointestinal EIA

Adenovirus: Collect fresh stool in sterile, leakproof container without media, serum, preservative, or metal ion. For patients requiring the use of diapers, first line the diaper with clean plastic to prevent absorption. Then, transfer 2 g or 2 mL of the stool specimen from the plastic-lined diaper to the sterile container. Do not submit the diaper itself. Cap securely. Do not use VCM or equivalent. Do not use any preservative, media, or additive. Swabs are not acceptable. Store refrigerated. Stable up to 3 days. Store frozen for longer stability.

Rotavirus: Collect stool specimens as for Adenovirus EIA. Do not use VCM or equivalent. Swabs are acceptable if not placed in VCM or equivalent. Store refrigerated. Stable up to 3 days. Store frozen for longer stability.

All viral cultures

“Source” information is required by CAP and federal regulations. The source or anatomic site of the specimen submitted for culture determines the type(s) of virus isolated. Clinicians may also indicate specific virus(es) to include in the culture. The following are routinely isolated: Adenovirus, Cytomegalovirus, Enterovirus, Herpes simplex virus, Influenza A & B, Parainfluenza 1, 2, & 3, Respiratory syncytial virus, Varicella/Herpes zoster.

The following are non-routine, requiring specific request: Measles and Mumps. It is preferable to store and immediately transport specimens refrigerated. If delay is unavoidable and the specimen is collected in VCM or equivalent transport medium, storing and transporting at -70°C (dry ice) is acceptable for almost all viruses. If the transport medium used is not VCM or equivalent, do not freeze specimens for Varicella/Herpes zoster, RSV, Measles, or CMV. Whole blood or bone marrow are NOT transported in VCM or equivalent and cannot be frozen (see below).

Biopsy/autopsy tissue

Obtain each specimen with a separate set of sterile instruments and place in separate tubes of VCM or equivalent. If VCM or equivalent is not available, place in separate sterile containers and add TSB. At the very least, use sterile normal saline. Do not add fixative. Tissues in VCM or equivalent may be frozen (-70°C/dry ice) for up to 5 days.

Blood

Collect in sodium heparin (adults 7.5-10 mL whole blood, pediatrics 3-7.5 mL). Do not use VCM or equivalent. Do not freeze. Note: for CMV, do not use culture; Collect whole blood in EDTA for PCR.

Bone marrow

Collect 0.3 mL of aspirate or core biopsy of approximately 1.5 x 0.3 cm size. VCM - Viral-Chlamydial- Mycoplasma transport medium (green-cap) available from client supplies. Cap tightly.

Bronchoalveolar lavage

Wedge bronchoscope into subsegmental bronchus; insert four 50 mL boluses of sterile saline into the suction port with immediate return suction after the insertion of each sample. Submit 5-10 mL in a sterile container.

Cervix

Remove mucous with sterile swab and discard. Use fresh sterile swab to obtain cellular material. Use VCM or equivalent. May freeze at -70°C (dry ice) for longer stability.

Conjunctiva

Use sterile swab to remove any exudate or mucous, then discard. Use normal sterile saline to pre-moisten a sterile swab, then firmly rub over the palpebral conjunctiva. Use VCM or equivalent. May freeze at -70°C (dry ice) for longer stability.

CSF

Collect aseptically. Submit 1-2 mL in sterile leakproof polypropylene container. Or add in equal proportion to VCM or equivalent (i.e., I mL CSF + I mL VCM or equivalent). May freeze at -70°C (dry ice) for longer stability, only if in VCM or equivalent.

Endocervical swab

Use a dacron/rayon swab or cleansing swab provided to remove mucus and exudate from the endocervix. Insert the swab into the endocervix, rotate, and remove. Discard. Insert a second swab into the cervical os to collect cells from the transitional zone. Rotate the swab for 10-30 seconds in firm contact with endocervical surfaces. Withdraw the swab without touching any vaginal surfaces. Place the swab in the Microtest (VCM) microbe transport tube (blue or red cap or equivalent), break off the excess, and cap tightly.

Endourethral swab

Patient should not have urinated for at least one hour. Insert a small wire-shafted dacron swab 2-4 cm into the endourethra. Gently rotate the swab. Wait 2-3 seconds. Withdraw the swab. Place the swab in the Microtest (VCM) microbe transport tube (blue or red cap or equivalent), break off the excess, and cap tightly. 

Fluids (bronchial, lavage, nasal, aspirate, sputum, pleural, peritoneal, etc.)

Collect aseptically. Submit in equal proportion to VCM or equivalent (i.e., 3 mL fluid + 3 mL VCM or equivalent). Do not freeze unless added to VCM or equivalent.

Lesion (vesicular)

Open the lesion, then use sterile swabs to collect fluid and cells from base of lesion. Use VCM or equivalent. May freeze at -70°C (dry ice) up to 5 days only if in VCM or equivalent.

Nasal aspirate

Use a suction apparatus or 5 cc syringe attached to a number 8 French catheter threaded 1-2 cm into the anterior nares to remove mucus, cells, and nasal fluid. Expel the aspirate into a sterile container.

Nasal wash

Use a sterile disposable pediatric ear syringe bulb containing 3-5 mL of sterile physiologic saline. For a child or infant, place the patient on his side. Gently press the upper nostril closed with finger pressure. Insert the tip of the syringe bulb into the lower nostril. Inject the saline into the open nostril and immediately aspirate the saline back into the bulb with a squeeze-release action. For an adult, have the patient close the epiglottis and tilt the head back. Instill the saline into each nostril. Collect the saline into a sterile container as the patient brings his head forward.

Nasal turbinate swab

Vigorously rotate 2 dacron swabs against the nasal turbinate bilaterally. Place the swabs into a VCM - Viral-Chlamydial-Mycoplasma transport medium (green-cap) available from client supplies, break off the excess, and cap tightly.

Nasopharyngeal

Insert flexible fine-shafted sterile swab into nostril to the posterior nasopharynx and gently rotate. Use a second sterile swab for the other nostril. Combine both into one tube of VCM or equivalent. May also combine with throat swab in one tube of VCM or equivalent. If in VCM or equivalent, may freeze at 70°C (dry ice) up to 5 days.

Rectal swab

Insert a dacron swab into the anal orifice 3-5 cm past the anal sphincter. Rotate the swab and withdraw. Place the swab in the VCM - Viral-Chlamydial-Mycoplasma transport medium (green-cap) available from client supplies; break off the excess. Cap tightly.

Sputum

Collect expectorate in response to a deep cough and place in a sterile container.

Stool

Collect feces in clean, dry container. Submit several grams undiluted in sterile leakproof container. Do not add fixative or preservative. May freeze at -70°C (dry ice) up to 5 days if in VCM or equivalent. E IA: Submit in a stool container - do not place stool into VCM - Viral-Chlamydial-Mycoplasma transport medium (green-cap) available from client supplies.

Throat

Rub one or two sterile swabs (pre-moistened with sterile normal saline) over the posterior wall of the pharynx. Use VCM or equivalent. May freeze at -70°C up to 5 days if in VCM or equivalent.

Throat wash

Have the patient gargle with 3-5 mL of sterile physiologic saline. Collect the saline in a sterile container.

Urine

Obtain fresh clean-catch urine in sterile leakproof container. Submit 2-10 mL. If VCM or equivalent is available, urine may be added in equal proportion (i.e. 3 mL urine + 3 mL VCM or equivalent). Do not freeze unless in VCM or equivalent.

Vesicle

Rupture the vesicle. Using a dacron swab, rub the fluid and cells from the base of the vesicle. Place the swab in a VCM - Viral-Chlamydial-Mycoplasma transport medium (green-cap) available from client supplies. Break off the excess and cap tightly. Sample several early stage cutaneous lesions if possible. Do not use local disinfection until after specimen collection.

Frozen samples

Frozen samples must be submitted in plastic containers. Do not submit frozen samples in glass containers. If multiple tests are requested on frozen samples, submit a separate frozen sample for each test ordered so that each analyte is properly preserved. Do not freeze samples for viral culture at regular freezer temperature. Freeze at -70° C or lower.

The following are non-routine, requiring specific request: Measles and Mumps. It is preferable to store and immediately transport specimens refrigerated. If delay is unavoidable and the specimen is collected in VCM or equivalent transport medium, storing and transporting at -70˚C (dry ice) is acceptable for almost all viruses. If the transport medium used is not VCM or equivalent, do not freeze specimens for Varicella/Herpes zoster, RSV, Measles, or CMV. Whole blood or bone marrow are NOT transported in VCM or equivalent and cannot be frozen.

Acute/convalescent paired sera

When both acute and convalescent samples are identified and sent together, they will be run in parallel and the base fee for two assays will be charged.

Acute samples received earlier than their convalescent samples will be assayed for the single-test fee. A reduced fee of one-half of the base fee will be charged for repeating the acute assay in parallel with the convalescent sample if, when received, the convalescent sample contains the acute sample’s identification number.

Repeat determinations

Repeat determinations are performed at NO CHARGE whenever reported results do not, in the opinion of the physician, fit the clinical picture of the patient. If a repeat of the original test is desired, please contact the lab promptly before the sample is discarded.

Packaging STAT specimens

Put the STAT specimen in a plastic specimen bag and place a STAT sticker on the bag. If the specimen is being transported frozen, tape the STAT label to the bag so that the label does not fall off when the bag is placed on dry ice. Mark the STAT box on the Request Form and indicate the telephone number to call with results. Place the additional STAT sticker on top of the outermost bag or box, including any express carrier packaging.

Medicolegal clinical specimens

Continuity of possession must be documented in all phases of medicolegal specimen handling from the initial collection to the rendering of a report. A chain-of-custody form should be used to document the possession of such specimens. Chain-of-custody forms and other medicolegal supplies are available by contacting the Client Supplies Department.

Specimens should be sealed with security tape and labeled at the time of collection. The chain-of-custody form, which is a record of all persons having possession of the sample, should be started and a Request Form should be filled out with requested tests. Please be specific in your request and in the information you present. This will enable us to handle your specimen properly and thus provide you better service.

NOTE: An additional fee is required for this service in addition to the testing fee. If this is an autopsy specimen, clearly indicate on the request form next to the patient name.