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Lab Manual for UCSF Clinical Laboratories

Lab Manual for SFGH

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VIROLOGY, ROUTINE CULTURE AND OTHER DETECTION METHODS

Item Value
Approval req'd? yes, see specific test
Available Stat? No
Test code VIRC
Performed by? Microbiology
Sendout? yes, see specific test
Price range $$$
In House Availability 7 days
Principle Specimens are inoculated to appropriate cell lines, which are incubated. Cell lines are examined daily for appearance of cytopathic effect (CPE). Identity of viruses is made through recognition of typical CPE and confirmation by appropriate fluorescent antibody staining. Cultures are held for 2 weeks before being reported as "Negative."

Specimens for detection of cytomegalovirus are inoculated to appropriate cells in shell vials and stained for CMV early antigen after 24 and 48 hours incubation.
Interpretation INTERPRETATION: Positive result indicates the presence of virus in culture. Negative result (no evidence of viral infection after 2 weeks incubation) may indicate absence of virus or may be due to improper specimen collection, improper specimen handling, or small amounts of virus which were not detected.

CLINICAL SIGNIFICANCE: The result must be interpreted in conjunction with clinical findings and other laboratory data.
Container type For collecting specimens which require a use of a swab, use swabs packaged with viral transport medium (obtained from Lab Support Services 206-8199). If necessary, swabs may be slightly moistened with sterile saline before collecting the specimen. (DO NOT use the viral transport medium to premoisten swab.) After collecting specimen, aseptically break swab into tube of transport medium and cap tightly. If viral transport packages are not available, dacron swabs with plastic or fine-wire shafts may be used and placed into a small amount (approx. 0.5 mL) of nonbacteriostatic saline in a capped sterile tube for transport to the lab. Dry swabs will not be processed. DO NOT USE CALCIUM ALGINATE SWABS as they may inactivate Herpes viruses.

Put other specimens for viral culture in sterile, leak-proof containers.
Collection Instructions Collect specimens aseptically as soon as possible after onset of symptoms or at autopsy. Recovery of viruses is best during the first 3 days after onset and may be greatly reduced after 5 days.

Blood
Blood must be collected in a lavender top (EDTA) tube. Green top (Heparin) tube is NOT acceptable. Culture of blood is not performed. Detection of virus is blood is done by PCR. See ARUP web page for Cytomegalovirus by PCR, Whole Blood or Bone Marrow test details..

Bone marrow
Blood must be collected in a lavender top (EDTA) tube. Green top (Heparin) tube is NOT acceptable. Culture of bone marrow is not performed. Detection of virus in bone marrow is done by PCR. See ARUP web page for Cytomegalovirus by PCR, Whole Blood or Bone Marrow test details..

Bronchoalveolar Lavage (BAL)
Collect 8-10 mL in sterile, screw-capped container.

Cerebrospinal fluid
Viral cultures are not performed on CSF. Please contact the Microbiology Laboratory Medicine Resident (206-5699 or pager 415-443-1438), Monday through Friday between 9:00 am and 5:00 pm for more information or to discuss alternative tests.

Conjunctiva
Use small-tip dacron swab packaged with viral transport medium. Slightly moisten swab with sterile saline and swab lower conjunctiva. After collecting specimen, aseptically break swab into tube of transport medium and cap tightly.

Lesions See Vesicles and Skin Scrapings

Nasal swabs
Use swab packaged with viral transport medium. Rotate swab in nostril and let it rest for several seconds to absorb secretions. After collecting specimen, aseptically break swab into tube of transport medium and cap tightly.

Nasopharyngeal aspirate/wash
(Preferred specimen for isolation of respiratory viruses)
Suction nasopharyngeal secretions directly into collection device. Add a small amount (< 1mL) of non-bacteriostatic saline to keep specimen moist. Or introduce approx. 2 mL sterile saline into nasopharynx and apply suction to obtain specimen. For respiratory syncytial virus or influenza A and B viruses, see also Respiratory Syncytial Virus (RSV), Rapid Detection .

Nasopharyngeal swab
Use small-tip dacron swab (on flexible wire shaft) packaged with viral transport medium. Insert swab through a nostril and into the posterior nasopharynx and rotate the swab several times to collect mucosal epithelium. After collecting specimen, aseptically break swab into tube of transport medium and cap tightly.

Pericardial fluid
Collect at least 2 mL in sterile screw-capped container.

Peritoneal fluid
Collect at least 2 mL in sterile screw-capped container.

Pleural fluid
Collect at least 2 mL in sterile screw-capped container.


Rectal swab
Use swab packaged with viral transport medium. Insert swab 4-6 cm into rectum and roll against mucosa. After collecting specimen, aseptically break swab into tube of transport medium and cap tightly.

Sputum
Collect 1-5 mL in sterile, screw-capped container.

Stool
Collect 1 - 2 teaspoonsful per specimen. Send specimen in screw-capped plastic container. Stool specimens will not be accepted for CMV Culture.

Throat swab
Use swab packaged with viral transport medium. Rub tonsillar or posterior nasopharynx area vigorously. After collecting specimen, aseptically break swab into tube of transport medium and cap tightly.

Tissue
Collect tissues from probable sites of pathology using a separate sterile instrument for each sample. Samples should be about 1 cm3. Place each specimen in a separate sterile container, keep chilled at 4°C, and deliver to Microbiology Laboratory (2M33) immediately. A small amount (approx. 0.5 mL) of non-bacteriostatic saline may be added to the specimen to prevent drying.

Ulcers
See Vesicles and Skin Scrapings

Urine
Collect 10-20 mL of midstream, clean-voided urine in sterile, screw-capped container.

Vesicles and Skin Scrapings (lesions, ulcers)
Use swab provided with viral transport medium to absorb vesicular fluid and scrape or rub vigorously the base of lesion. It is essential to obtain cells (parabasal layer). For ulcers, gently wash away necrotic debris before swabbing. After collecting specimen, aseptically break swab into tube of transport medium and cap tightly.

Do not use calcium alginate swabs as calcium alginate may inactivate herpes viruses.


Special instructions REQUISITION: Microbiology (Viral - Routine Culture, Routine viral culture [NOT including CMV], or Routine viral culture including CMV).

Please send properly completed form, noting date and time of collection, source of specimen, test requested, and suspected virus. SOURCE OF SPECIMEN IS REQUIRED.

Deliver to Microbiology Lab (2M33) immediately or hold at 4° C until delivery.
Normal range no virus recovered
Synonyms Adenovirus Isolation;Autopsy Viral Culture;BAL (Bronchoalveolar Lavage) Culture;Biopsy Culture;Bronchoalveolar Lavage (BAL) Culture;Coxsackie Virus Isolation;Echovirus Isolation;Enterovirus (Coxsackievirus, Echovirus, Poliovirus) Isolation;Influenza Virus Isolation;Lesion, Viral Culture;Nasopharyngeal Aspirate, Viral Detection From;Parainfluenza Virus Isolation;Pericardial Fluid Culture;Peritoneal Fluid Culture;Pleural Fluid Culture;Polio Virus Isolation;Respiratory Virus Isolation;Stool Culture;Throat Culture;Ulcer Culture;Urine Culture;Varicella-Zoster Virus Isolation;Virology, Collection and Transport of Specimens;VZV;Respiratory syncytial virus (RSV) isolation;
Stability The stability of viruses varies from relatively labile (CMV, varicella-zoster) to relatively stable (enteroviruses). For optimal isolation of viruses, hand carry specimens to the Microbiology Lab (2M33) directly after the specimen is collected. Specimens delivered by messenger service may be delayed in transit, and isolation of viruses may thus be compromised. If specimens are not delivered immediately, store at 4°C. NEVER freeze specimens in household-type refrigerator/freezer (-20°C). This will severely decrease viral infectivity. All specimens must be received in the lab within 24 hours of collection.
Additional information Routine viral culture includes appropriate culture for adenoviruses, enteroviruses (coxsackie, echo and polioviruses), Herpes simplex virus Types 1 and 2; influenza viruses, parainfluenza viruses, and Varicella-zoster virus. Please indicate suspected virus on requisition.

Because methods for optimal detection of cytomegalovirus may vary from methods for detection of the above viruses, cytomegalovirus must be specifically requested. See Cytomegalovirus (CMV) Isolation.


References 1. Clarke, LM, ed. 1992. Section 8: Viruses, Rickettsiae, Chlamydiae, and Mycoplasmas; as found in Clinical Microbiology Procedures Manual, H. Isenberg, ed. Amer. Soc. for Microbiol., Washington, D.C.

2. Schmidt, NJ and RW Emmons, eds. 1989. Diagnostic Procedures for Viral, Rickettsial and Chlamydial Infections, 6th ed. Amer. Public Health Assn, Washington, D.C.
CPT coding 87252
Last Updated 12/26/2013 11:11:11 AM
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