Clinics and Departments

Laboratory Services

Lab Dept:

Microbiology/Virology

Test Name:

VIRAL CULTURE

General Information

Lab Order Codes:

VIRC

Synonyms:

Viral Isolation; Virus Isolation; Virus Culture

CPT Codes:

87252 – Virus isolation; tissue culture inoculation, observation, and presumptive identification by cytopathic effect

The following testing may be added if appropriate based on findings for organism identification (multiple additions are possible if more than one organism is identified).

87253 – Virus isolation; tissue culture, additional studies or definitive identification, each isolate (if appropriate)

Test Includes:

The detection of adenovirus, cytomegalovirus, coxsackievirus, echovirus, enterovirus, herpes simplex virus, influenza virus, measles, mumps, parainfluenza virus, poliovirus, respiratory syncytial virus, rhinovirus, and varicella-zoster virus.

Logistics

Lab Testing Sections:

Virology

Phone Numbers:

MIN Lab: 612-813-5806

STP Lab: 651-220-6555

Test Availability:

Daily, 24 hours

Turnaround Time:

14 – 21 days

Special Instructions:

● Indicate the virus suspected.
● Requisition must state specific site of specimen and date/time of collection.
● Collect specimens early in the course of illness to yield highest viral titers.
Do Not use calcium alginate swabs

Specimen

Specimen Type:

Whole blood, cerebrospinal fluid, dermal, ocular, genital, mucosal, respiratory, oral, stool, rectal, urine, tissue, biopsy. See table in Additional Information.

Container:

Swab transport system, sterile container, viral transport media (M4 VTM)

Volume:

Blood:

1 – 5 mL in lavender top (EDTA) tube, See Viral Blood Culture

Bone Marrow:

1 – 5 mL in Lavender top (EDTA) tube

Bronchoscopy washings:

1 – 2 mL

Cervical:

1 swab

CSF:

1 mL

Dermal specimens:

1 swab

Genital specimens:

1 swab

Nasopharyngeal aspirates:

1 – 2 mL

Nasopharyngeal swabs:

2 NP swabs

Ocular/Conjunctiva specimens:

1 swab

Oral specimens:

1 swab

Rectal specimens:

1 swab

Stool:

1 gm solid stool/ 1.0 mL liquid stool

Throat specimens:

1 swab

Tissue:

Submit as much tissue as possible

Vaginal:

1 swab

Washings/Aspirates:

1 – 2 mL

Collection:

Blood: See Viral Blood Culture

Bone Marrow:

Place 1 – 5 mL of bone marrow in a lavender-top (EDTA) tube(s). Invert several times to mix bone marrow. Do Not centrifuge. Send in original Vacutainer tube. Forward unprocessed bone marrow promptly at ambient temperature only.

Bronchoscopy:

1. Collect 1 – 2 mL of specimen obtained by physician through the biopsy channel of the bronchoscope.
2.
Transfer specimen into a luki tube.
3.
Transport to the Microbiology Laboratory immediately.

Cervical:

1. If lesions are present, swab vigorously with a sterile swab.
2.
If lesions are not present, remove exudates prior to collection of specimen.
3.
Gently insert separate large swab into endocervical canal past squamocolumnar junction (1 cm into the cervical canal). Rotate for 5-10 seconds.
4.
To avoid contamination, withdraw the swab while avoiding touching any vaginal surfaces.

CSF:

1. Disinfect skin site with 2% tincture of iodine.
2.
Insert needle with stylet at L3 – L4, L4 – L5, or L5 – S1 interspace.
3.
Upon reaching the subarachnoid space, remove the stylet and collect 1 – 2 mL of fluid into each of 3 sterile CSF tubes.
4.
Deliver tube #2 to Microbiology Lab immediately.
5.
Do Not place specimen in viral transport media.

Dermal/Skin:

1. Wash vesicles with sterile saline.
2.
Unroof the vesicle and absorb vesicular fluid into a dry swab.
3.
Vigorously scrape base of freshly exposed lesion with swab to obtain cells which contain virus.

Nasopharyngeal aspirates:

1. Prepare suction set up on low to medium suction.
2.
Wash hands and put on protective barriers (e.g., gloves, gown, mask)
3.
Place child supine and obtain assistant to hold child during procedure.
4.
Attach luki tube to suction tubing and #6 French suction catheter.
5.
Insert catheter into nostril and pharynx without applying suction.
6.
Apply suction as catheter is withdrawn.

Nasophyngeal swabs:

1. Obtain 2 specimens using 2 NP swabs (i.e., MiniTip™ Culturette).
2.
Gently insert swab through nose into posterior nasopharynx.
3.
Gently rotate swab slowly for 5 seconds to absorb secretions.
4.
Collect a second specimen in the same manner.

Ocular/Conjunctiva:

Do Not use a dry swab to collect an eye culture.

1.
Moisten swab with sterile saline.
2.
Retract lower lid and firmly swab conjunctival surface with enough pressure to collect epithelial cells. Avoid eyelid border and lashes.

Oral:

1. Firmly sample base of lesion(s) with swab.

Rectal:

1. Insert swab approximately 1 inch into anal canal.
2.
Gently move the swab from side to side to sample the anal crypts.

Stool:

1. Collect stool in a clean, dry bedpan or on a newspaper over the toilet.
2.
Transfer specimen to a plastic, leakproof container. Do not overfill or contaminate the outside of the container.
3.
Forward promptly to Microbiology Lab and refrigerate.

Throat:

1. Depress the tongue with a tongue depressor so the swab does not touch the tongue.
2.
Sample the posterior pharynx, tonsils, and inflamed areas with a sterile swab.

Tissue:

1. Submit specimen in a screw capped, sterile container.
2.
Maintain sterility and forward promptly.

Urine:

Males:

1. Clean glans with soap and water.
2.
Rinse area with wet gauze pads.
3.
While holding foreskin retracted, begin voiding.
4.
After several mL have passed, collect a minimum of 5.0 mL without stopping flow of urine.
5.
Maintain sterility and forward immediately to the Microbiology Lab. Refrigerate.

Females:

1. Thoroughly clean urethral area with soap and water.
2.
Rinse area with wet gauze pads.
3.
While holding labia apart, begin voiding.
4.
After several mL have passed, collect a minimum of 5.0 mL without stopping flow of urine.
5.
Maintain sterility and forward immediately to the Microbiology Lab. Refrigerate.

Vaginal:

1. Wipe away excessive amount of secretion or discharge.
2.
If lesions are present swab vigorously with a sterile swab.
3.
If lesions are not present, obtain secretions from mucosal membrane of the vaginal vault with a sterile swab.

Special Processing:

Extract swab specimen thoroughly into transport medium by swirling and pressing swab against the inside of the vial, then discard swab. Allow NP swabs to remain in the media by cutting the shaft of the swab. Cap tightly.

Lab Staff:

Urine: Add 3 - 5 mL urine to urine VTM. Refrigerate.

Washings/Aspirates: Place washings/aspirates into VTM. Refrigerate.

Tissue: Place tissue into VTM. Refrigerate.

Stool: Place 1 mL of liquid stool or a pea size aliquot of formed stool into a thawed 9 mL VTM to make a 1:10 dilution. Vortex well and refrigerate.

Transport/Storage:

● Maintain sterility and transport to the Microbiology Laboratory immediately. Store at refrigerated temperatures. Do Not freeze.
● If there is a delay in transport of 1 hour or more, place swab, washings/aspirate and tissue specimens in viral transport media and refrigerate.

Sample Rejection:

Specimen with a transit time exceeding 2 hours after collection; specimen not submitted in appropriate transport container; improperly labeled specimen; insufficient volume; external contamination. If an unacceptable specimen is received, the physician or nursing station will be notified and another specimen will be requested before the specimen is discarded.

Interpretive

Reference Range:

No Virus isolated

Critical Values:

Positive results in systemic infections will be called to the physician or nursing unit.

Limitations:

● A negative result does not eliminate the possibility of viral infection.
● HSV can only rarely be cultured from the CSF. HSV PCR is the method of choice for detecting HSV in CSF specimens.
● Isolation of a virus may not be related to the patient’s disease.

Methodology:

Conventional cell culture with specific confirmation procedures of viral agent.

Additional information:

Viruses Typically Isolated From Clinical Specimens

Specimen Type

Virus

Blood

CMV (Enterovirus#, HSV#, VZV#)

CSF and CNS Tissues

Enterovirus, Mumps#, HSV#, CMV#

Dermal Lesions

HSV, VZV, Adenovirus, Enterovirus, Rubeola

Eye

HSV, VZV, Adenovirus, Enterovirus, CMV

Genital

HSV, CMV

Oral

HSV, VZV#, Enterovirus

Respiratory Tract

Upper

Lower

 

Adenovirus, Rhinovirus, Influenza, HSV, Parainfluenza, Enterovirus, RSV

Adenovirus, Influenza, Parainfluenza, RSV, CMV

Stool

Enterovirus, Adenovirus

Tissues

CMV, HSV, Enterovirus, Adenovirus

Urine

CMV, Adenovirus, Enterovirus, Mumps

Abbreviation Key

HSV

Herpes Simplex Virus

CMV

Cytomegalovirus

VZV

Varicella-Zoster Virus

RSV

Respiratory Syncytial Virus

Enteroviruses include:

Coxsackie Virus, Poliovirus, Echovirus, Enterovirus

#

Indicates virus specified is less frequently isolated.

References:

Cook, JH, and M Pezzlo (1992). Specimen receipt and accessioning. Section 1. Aerobic bacteriology, 1.2.1-4. In HD Isenberg (ed) Clinical Microbiology Procedures Handbook. American Society for Microbiology, Washington DC

Miller, J Michael (1999) A Guide To Specimen Management in Clinical Microbiology, American Society for Microbiology, Washington DC

Miller, J Michael, and HT Holmes (1999) Specimen Collection, Transport, and Storage In PR Murray et al, (ed), Manual of Clinical Microbiology, 7th edition, American Society for Microbiology, Washington DC, pg 33-104

Updates:

3/25/2010: CPT Updates



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