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

Molecular detection and genotyping of dengue (DEN) 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 cerebrospinal 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 dengue virus infection and relevant travel history. Reactive dengue virus 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. Johnson, B.W., Russell, B.J. and Lanciotti, R.S. Serotype-specific detection of dengue viruses in a fourplex real-time reverse transcriptase PCR assay. 2005. J. Clin. Micro. 43; 4977-4983.
2. Lanciotti, R.S., Calisher, C.H., Gubler, D.J., Chang, G.J., and Vorndam, V. Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction. 1992. J. Clin. Micro. 30; 545-551.