Molecular Detection of Japanese Encephalitis Virus by Reverse Transcriptase PCR (RT-PCR)

Molecular detection of Japanese encephalitis (JE) virus by reverse transcriptase PCR. Amplicons generated are subjected to DNA sequencing.

Plasma is the preferred specimen; however testing can be performed on serum or cerebral spinal fluid (CSF). In cases of central nervous system infection, CSF can be tested to demonstrate that the virus has crossed the blood-brain barrier. Minimum volume of 400 uL.

Collect blood in tubes with an anticoagulant (not heparin).

Store samples frozen until shipped for testing. Ship samples on dry ice.

Shipping of specimens shall be done by a TDG certified individual in accordance with TDG regulations. For additional information regarding classification of specimens for the purposes of shipping, consult either Part 2 Appendix 3 of the TDG Regulations or section 3.6.2 of the IATA Dangerous Goods Regulations as applicable.

Suspected JE virus infection and relevant travel history. Reactive JE serology.

Completed Viral Zoonoses requisition including sender laboratory name, address and telephone number. Patient name and / or identifier (specimen reference number), date of birth, test(s) requested, collection date of specimen, date of on-set of symptoms, type of specimen, and clinical and travel history of patient.

This is not a routine test. Please contact the Viral Zoonoses Diagnostic Laboratory before sending specimens.

Reverse-transcriptase PCR. Identification of amplicons by direct sequencing. Isolation of an arbovirus, or detection of arboviral antigen or nucleic acid in a clinical specimen would constitute firm evidence of viral association with illness and provide "confirmed case" status.

Please contact the laboratory for the turnaround time.

1. Ayers, M., Adachi, D., Johnson, G., Andonova, M., Drebot, M., and Tellier, R. A single tube RT-PCR assay for the detection of mosquito-borne flaviviruses. J Virol Meth 2006. 135:m 235-239.