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Test ID WNVUR West Nile Virus, RNA, PCR, Molecular Detection, Random, Urine


Specimen Required


Container/Tube: Sterile container

Specimen Volume: 1 mL

Collection Instructions:

1. Collect random urine in a sterile container.

2. No preservative should be added.

3. Label specimen as urine.


Useful For

Rapid testing for West Nile virus (WNV) RNA (lineage 1 and lineage 2)

 

An adjunctive test to serology for detection of early WNV infection (ie, first few days after symptom onset), with urine specimens potentially retaining WNV RNA longer than other sources

 

This test should not be used for screening asymptomatic individuals and should only be used to test patients with signs and symptoms of WNV disease.

Method Name

Real-Time Reverse Transcription Polymerase Chain Reaction (RT-PCR)

Reporting Name

West Nile Virus RNA, PCR, Urine

Specimen Type

Urine

Specimen Minimum Volume

0.3 mL

Specimen Stability Information

Specimen Type Temperature Time Special Container
Urine Refrigerated (preferred) 7 days
  Frozen  7 days

Reject Due To

Urine containing preservatives Reject
Heat-inactivated specimen Reject

Clinical Information

West Nile virus (WNV) is a mosquito-borne flavivirus (single-stranded RNA virus) that primarily infects birds but occasionally infects horses and humans.(1,2,3) Until the virus was recognized in 1999 in infected birds in New York City, WNV had been detected only in the Eastern hemisphere, with a wide distribution in Africa, Asia, the Middle East, and Europe. There are 2 distinct lineages of WNV: lineage 1 has the broadest distribution worldwide, including North America and Europe, whereas lineage 2 is found only in Africa and parts of Europe.

 

Most people who are infected with WNV do not experience symptoms. It is estimated that about 20% of those who become infected will develop West Nile fever with mild symptoms, including headache, myalgia, and, occasionally, a skin rash on the trunk of the body. About 1 of 150 WNV infections (<1%) results in meningitis or encephalitis. Fatality rates among patients hospitalized during recent outbreaks have ranged from 4% to 14%. Advanced age is the most important risk factor for death, and patients older than 70 years are at particularly high risk.

 

Laboratory diagnosis is best achieved by demonstration of specific IgG- and IgM-class antibodies in serum specimens. However, polymerase chain reaction (PCR) testing can be used to detect WNV RNA in serum, whole blood, and urine specimens from patients with recent WNV infection (ie, 3-5 days following infection) when specific antibodies to the virus are not yet present. It may also be useful for patients who are immunocompromised when an antibody response is minimal or absent. Finally, PCR can be useful for supporting a serologic diagnosis, given the known cross-reactivity of WNV serology with other flaviviruses.

 

Studies indicate that whole blood testing by PCR may provide higher sensitivity when testing patients with acute WNV disease (up to 87%) compared to serum, plasma, urine, and cerebrospinal fluid testing.(4) However, viral RNA may be detected for a longer period of time (≥10 days after symptom onset) in urine than in other sources.(5) Serum testing offers lower sensitivity (26%) but may be used when it is the only specimen type available.

Reference Values

Negative

Reference values apply to all ages.

Interpretation

A positive result indicates the presence of West Nile virus RNA and is consistent with early infection.

Cautions

The sensitivity of the assay is very dependent upon the time of illness onset in which the specimen is collected. Polymerase chain reaction testing has the greatest utility when used within the first few days of symptom onset.

 

A negative test does not exclude infection with West Nile virus. Therefore, the results obtained should be used in conjunction with clinical findings and serologic test results to make an accurate diagnosis.

 

This assay detects both viable and nonviable virus. Test performance depends on viral load in the specimen and may not correlate with cell culture performed on the same specimen.

Clinical Reference

1. Petersen LR, Brault AC, Nasci RS. West Nile virus: Review of the literature. JAMA. 2013;310(3):308-315

2. Colpitts TM, Conway MJ, Montgomery RR, Fikrig E. West Nile virus: biology, transmission, and human infection. Clin Microbiol Rev. 2012;25(4):635-648

3. Centers for Disease Control and Prevention (CDC), National Center for Emerging and Zoonotic Infectious Disease (NCEZID), Division of Vector-Borne Diseases (DVBD): West Nile Virus: Information for Healthcare Providers. CDC; Reviewed April 25, 2023. Accessed February 6, 2024. Available at www.cdc.gov/westnile/healthcareproviders/index.html

4. Lustig Y, Mannasse B, Koren R, et al. Superiority of West Nile virus RNA detection in whole blood for diagnosis of acute infection. J Clin Microbiol. 2016;54(9):2294-2297

5. Barzon L, Pacenti M, Franchin E, et al. Excretion of West Nile virus in urine during acute infection. J Infect Dis. 2013;208(7):1086-1092

Method Description

This real-time reverse transcription polymerase chain reaction (RT-PCR) assay provides qualitative detection of lineages 1 and 2 of West Nile virus. Viral RNA is first extracted from urine using an extraction platform. The extract is then used for subsequent RT-PCR. An initial reverse transcription step is employed to convert viral RNA to complementary DNA (cDNA). The viral cDNA target is then amplified on the Roche LightCycler 480 using specific primers and detected via Taqman probe technology. An internal control is also included with each reaction. Analysis of the PCR amplification is accomplished through the use of LightCycler software.(Package insert: RealStar WNV RT-PCR Kit 2.0. Altona Diagnostics; 03/2019)

Day(s) Performed

Monday through Friday

Report Available

Same day/1 to 5 days

Test Classification

This test was developed and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. It has not been cleared or approved by the US Food and Drug Administration.

CPT Code Information

87798

NY State Approved

Yes

Forms

If not ordering electronically, complete, print, and send a Microbiology Test Request (T244) with the specimen.