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Virus Isolation, Rapid Culture with Reflex

Updated

8/15/2013

Other Names
Virus Culture, Cell Culture, Respiratory Virus Culture, Enterovirus Culture, Varicella Culture, Respiratory Syncytial Virus (RSV), RSV
How to Order
Check the “Virus Isolation” box on the Virology/Immunology Request Form (form 42)
Availability
All submitters
Methodology
Spin Amplified Cell Culture with Reflex to Direct Fluorescent Antibody (DFA) Stain 
Performed
Monday through Friday
Time to Reporting
2 - 10 days after receipt of specimen
CPT Code
87254. 87140 x number of additional stains required to identify a positive.


Specimen Collection Details

Collection
  
Specimen
Common Viruses Isolated
CSF & CNS Tissues
CMV, HSV, enteroviruses, VZV
Dermal Lesions
Adenovirus, enteroviruses, HSV, VZV
Eye
CMV, Adenovirus, enteroviruses, HSV, VZV
Genital 
HSV
Mucosal 
HSV, VZV
Oral 
HSV, VZV
Rectal 
Enteroviruses, HSV, VZV
Respiratory Tract 
 
Upper 
Adenovirus, enteroviruses, HSV, hMPV, influenza, parainfluenza, rhinovirus, RSV
Lower 
Adenovirus, CMV, human metapneumovirus (hMPV), influenza, parainfluenza, RSV
Stool 
Adenovirus, enteroviruses
Tissues 
CMV, enteroviruses, HSV
Urine 
Adenovirus, CMV, enteroviruses
 
Cerebrospinal Fluid (CSF) - Do not use any transport media with CSF specimens. Place CSF in a sterile tube and seal tightly. Label with the client’s name and the date of collection. If the specimen will be held more than 1 week it should be frozen, preferably at  -70°C (-20°C if ultracold storage is not available), and shipped frozen to the OSPHL on cold packs or dry ice as soon as possible. Always ship CSF on a cold pack, regardless of holding time.
 
Other Body Fluids - Collect and ship as for CSF. Specimens may be placed in a small amount (one part viral transport media to four parts specimen) of viral transport media for stabilization, if desired. Blood and blood components are not acceptable body fluid specimens.
 
Stools - Stool should be collected in a sterile container that should be sealed tightly to prevent leakage and refrigerated, not frozen. Specimens should be shipped on a cold pack to OSPHL as soon as possible.
 
Swabs - Swab the throat, nasal cavity, vesicle, or other site thoroughly.  Place the swab in the M4 viral transport media tube. Break off the swab so that the cap can be screwed on the bottle securely. Do not remove the swab from the container before sending to OSPHL. Ship the specimen on a cold pack to OSPHL as soon as possible.
 
Urine - Collect about 10 ml clean-catch midstream urine in a sterile container and seal tightly. Refrigerate the urine specimen at 4°C and ship to the laboratory within 24 hours on cold packs. Do not freeze urines.
 
Tissues (Autopsy or Biopsy) - Collect tissue aseptically, using a separate set of instruments for each tissue and placing each in a separate sterile container containing viral transport media specify the site.  Send representative sections (preferably no larger than the size of a pea) of all infected organs. Ship specimens to OSPHL on cold packs as soon as possible.
 
Cell Culture Isolates for Identification - Select a cell culture tube or shell vial that exhibits 2+ or 3+ CPE (if possible), fill it with cell culture media, seal tightly, and ship to OSPHL as soon as possible. Cell cultures for identification must not be refrigerated or heated above 37°C at any time prior to their receipt by OSPHL.
Optimal Volume
2-3ml
Minimum Volume
1 ml
Handling
See collection
Transport
Transport specimens promptly, on ice or cold packs. DO NOT FREEZE.  Refrigerate if specimens must be held prior to shipment.
Rejection Criteria
- Blood or serum specimens.
- Specimens that have dried out (swabs, sputum, fluids, etc).
- Specimens collected using wooden shafted swabs.
- Specimens collected with calcium alginate or non-synthetic tipped swabs.
- Specimens that have been collected in non-sterile containers.
- Specimens collected in expired transport media.
- Specimens frozen at -20°C if sent for cytomegalovirus (CMV), respiratory syncytial virus (RSV), or varicella zoster virus (VZV).
- Specimens other than tissues sent in bacterial transport media (media which contains agar).
- Stool where isolation of CMV is requested.
- Preserved specimens (specimens in alcohol, formalin, etc.).
- Specimens collected more than 7 days after the onset of symptoms (certain specimens may be processed after consultation with the submitter so long as a comment is attached to the report).
- Specimens in leaky or broken containers.
- Specimens with insufficient quantity to perform the requested isolation with the exception of CSF (a comment will be added to the report).
- Unlabeled or incompletely labeled specimens
Comments
Specimens should be collected as early in the acute stage of infection as possible. Providing date of onset and a brief clinical description (e.g. if patient has rash or respiratory symptoms) can be helpful for virus culture.
     It is not practical to try to culture many common viruses that infect humans. Some examples are: hepatitis viruses, arboviruses, papillomaviruses, parvoviruses, reoviruses, and gastrointestinal viruses such as astro, corona, calici, and rotaviruses. 
     Respiratory cultures: Nasopharyngeal aspirates/washes or swabs are the preferred specimen types. Throat swabs, nasal aspirates, and bronchial washes may also be submitted. 
     Enterovirus cultures: Rectal swabs, stool, or Nasopharyngeal or lower respiratory tract specimens are acceptable. CSF, eye lesions, tissues, and urines will also be accepted.
     For other specimen types or suspected organisms, please consult with OSPHL before submission.