Molecular Detection of Zika Virus by Reverse Transcriptase PCR (RT-PCR)
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Requisition Forms
Reference Details
Molecular detection of Zika (ZIKV) virus by real-time reverse transcriptase PCR. Conventional RT-PCR may also be conducted if additional analysis is required.
- Zika virus infection
Serum* and urine* samples collected within 10 and 14 days respectively following symptom onset and as early/acute as possible once illness occurs. Additional samples for consideration for viral detection include CSF, placenta, amniotic fluid, and umbilical cord blood. Minimum volume of 600 µL required.
Isolation of virus may be attempted from some clinical specimens but is not deemed a routine procedure for confirming suspect cases of ZIKV infection.
Collect blood in serum separator tubes – submit in 2 ml O-ring tube. Collect urine in a sterile container – submit in 50 ml vial.
Store serum samples frozen until shipped for testing. Ship samples on dry ice. Store urine at 4°C and ship on wet ice for arrival at the NML within 48 hours of collection. Otherwise, freeze at -70°C and ship frozen 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 Zika virus infection and relevant travel history.
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.
Please note that the point of sample collection with respect to symptom onset is essential to help determine which tests are conducted and with interpretation of results. For specimens collected from female patients, please indicate whether the patient is pregnant or not.
Zika virus real-time RT-PCR based on TaqMan chemistry. Two separate singleplex assays are performed targeting two distinct genomic regions; a result is classified as positive if both targets are detected and each Ct value falls within a pre-established cut-off value for each assay. If only a single target is detected, the test is repeated and/or other testing platforms may be enlisted to resolve the interpretation of the result.
Turnaround time is 21 calendar days after date of receipt of the specimen. Samples requiring conventional PCR and sequencing have an extended turnaround time of 35 calendar days.
- Lanciotti, R.S., Kosoy, O.L., Laven, J.J., Velez, J.O., Lambert, A.J., Johnson, A.J., Stanfield, S.M., Duffy, M.R., (2008). Genetic and serologic properties of Zika virus associated with an epidemic, Yap State, Micronesia, 2007. Emerging Infectious Diseases, 14(8), 1232-1239