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Test Code POWVB Powassan Virus, RNA, Molecular Detection, PCR, Blood


Ordering Guidance


The Powassan virus polymerase chain reaction result may remain positive for a longer time in urine than in blood, serum, and cerebrospinal fluid (7 days or more).



Specimen Required


Container/Tube: Lavender top (EDTA)

Specimen Volume: 1 mL

Collection Instructions: Send whole blood specimen in original tube. Do not aliquot.


Secondary ID

617484

Useful For

Rapid testing for Powassan virus RNA (lineage 1 and lineage 2) using whole blood specimens

 

An adjunctive test to serology for detection of early Powassan virus infection (ie, first few days after symptom onset)

 

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

Testing Algorithm

For information see the following:

-Meningitis/Encephalitis Panel Algorithm

-Acute Tickborne Disease Testing Algorithm

Method Name

Real-Time Polymerase Chain Reaction (PCR)

Reporting Name

Powassan Virus, PCR, Blood

Specimen Type

Whole Blood EDTA

Specimen Minimum Volume

0.3 mL

Specimen Stability Information

Specimen Type Temperature Time Special Container
Whole Blood EDTA Refrigerated (preferred) 7 days
  Frozen  7 days

Reject Due To

  All specimens will be evaluated at Mayo Clinic Laboratories for test suitability.

Clinical Information

Powassan virus (POWV) is an emerging tick-borne virus, harbored by Ixodes species ticks, which are the same ticks that transmit Lyme disease (Borrelia spp.), Babesia spp., and Anaplasma phagocytophilum, among other pathogens. POWV is a member of the Flavivirus genus, which includes other arthropod-borne viruses (arboviruses) such as West Nile virus (WNV) and St. Louis encephalitis virus. Two lineages of POWV have been identified, sharing approximately 94% amino acid sequence identity, including lineage 1, which is the prototypical POWV lineage transmitted by Ixodes marxi and Ixodes cookei, and lineage 2, which includes deer tick virus and is transmitted by Ixodes scapularis. POWV is maintained in the environment in groundhogs, skunks, squirrels, and white footed mice. Unlike other tick-borne pathogens, following tick attachment to a host, POWV can be transmitted in as little as 15 minutes.

 

Following infection, the incubation period can last anywhere from 4 to 14 days, after which approximately 66% of patients will remain asymptomatic. Symptomatic patients may present with a nonspecific influenza-like illness, including high fever, fatigue, malaise, and myalgia. Approximately 30% of symptomatic patients will progress to develop neurologic manifestations, most commonly encephalitis. While some patients may recover, over 50% of individuals will have persistent neurologic sequelae. POWV has been associated with an overall mortality rate of 10%. Although there is no targeted antiviral therapy and treatment is entirely supportive care, diagnosis is important for a number of reasons, including the ability to discontinue empiric antibiotics and to provide prognostic information for patients and families.

 

While limited data exist for POWV real-time reverse transcription polymerase chain reaction (RT-PCR) detection, data from testing for the related flavivirus, WNV, suggests that blood may be a more sensitive source than serum for detecting viral RNA. Similarly, data from WNV and other flaviviruses suggest that viral RNA may be detected in urine for a longer period of time than in blood or serum. POWV RNA may be detected from cerebrospinal fluid in cases of neuroinvasive disease.

 

Powassan infections are often diagnosed based on a patient's symptoms and exposure risk in conjunction with molecular and serologic testing. The use of RT-PCR can provide a rapid laboratory confirmation of POWV RNA early in infection, particularly during the first 7 days of illness when serologic testing is typically negative. After 7 days, POWV RT-PCR is less sensitive and serologic testing is the preferred diagnostic method. It is common for RT-PCR and serology to be used together for diagnosis of early infection, as they are complementary methods.

Reference Values

Negative

Interpretation

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

Method Description

For this real-time reverse-transcription laboratory-developed polymerase chain reaction (PCR) assay, viral nucleic acid is extracted from specimens, followed by amplification and detection on the Roche LightCycler 480 instrument. This PCR assay has been optimized to detect a target sequence in the nonstructural protein. Primers amplify a 179 base pair product. Powassan virus genomic RNA is first transcribed to complementary DNA (cDNA) by reverse transcriptase, followed by amplification of the cDNA product. The LightCycler 480 is an automated instrument that amplifies and monitors the development of target nucleic acid (amplicon) after each cycle of PCR. The detection of amplicon is based on fluorescence resonance energy transfer, which utilizes a hybridization probe with a donor fluorophore, fluorescein, at the 3' end and a second hybridization probe with an acceptor fluorophore, LC-Red 610, at the 5' end. When the target amplicon is present, the LC-Red 610 emits a measurable and quantifiable light signal at a specific wavelength. Presence of the specific organism nucleic acid may be confirmed by performing a melting curve analysis of the amplicon. Using features of the melting curve analysis, the assay primers and specific hybridization probes are able to detect Powassan virus lineage 1 and lineage 2.(Unpublished Mayo method)

Day(s) Performed

Monday through Friday

Performing Laboratory

Mayo Clinic Laboratories in Rochester

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

LOINC Code Information

Test ID Test Order Name Order LOINC Value
POWVB Powassan Virus, PCR, Blood 34456-4

 

Result ID Test Result Name Result LOINC Value
617488 Lineage 1 34456-4
618144 Lineage 2 (Deer Tick Virus) 34456-4
618148 Powassan Virus PCR Comment 77202-0

Forms

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