Order Code POWVS Powassan Virus, RNA, Molecular Detection, PCR, Serum

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).

Additional Testing Requirements

This test is designed to be used during the first 7 days after symptom onset and should be ordered in conjunction with POWV / Powassan Virus, IgM, Enzyme-Linked Immunosorbent Assay, Serum.

Specimen Required

Collection Container/Tube:

Preferred: Serum gel

Acceptable: Red top

Submission Container: Sterile container

Specimen Volume: 0.5 mL

Collection Instructions:

1. Within 2 hours of collection centrifuge and aliquot serum into a sterile container.

2. Serum specimens not aliquoted from the serum gel collection tube into a sterile container will be rejected.

Useful For

Rapid testing for Powassan virus RNA (lineage 1 and lineage 2) using serum 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, Serum

Specimen Type

Serum

Specimen Minimum Volume

0.3 mL

Specimen Stability Information

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

Reject Due To

Gross hemolysis	Reject
Heat-inactivated specimen	Reject

Reference Values

Negative

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 179Â 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

CPT Code Information

87798

Forms

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

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