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Improved detection of dengue and Zika viruses using multiplex RT-qPCR assays

Abstract : Dengue virus (DENV) and Zika virus (ZIKV) are important viral pathogens, known to cause human infections with similar symptoms, are transmitted by common vectors and co-circulate in intertropical regions. Moreover, dengue fever results from infection with one of four different serotypes of dengue virus. Considering the recent ZIKV emergence, multiplex and up-to-date assays are more preferable for detection of both viruses in a single reaction. This study aimed to develop: (i) an one-step duplex real-time reverse transcription polymerase chain reaction (RT-qPCR) assay to efficiently and simultaneously detect and quantify DENV and ZIKV; (ii) a fourplex RT-qPCR to differentiate and quantify the four DENV serotypes. The detection limit of the duplex assay was 0.028 and 0.065 FFU (focus forming unit)/ml for DENV and ZIKV respectively. The lower limit of analytical sensitivity of fourplex assay was 0.01 FFU/ml for DENV-1 and 0.1 FFU/ml for DENV-2,-3 and-4. The assessment of specificity indicated both assays were highly specific to targeted viruses with negative results for other Flaviviridae such as Japanese encephalitis, West Nile, Yellow fever or Hepatitis C viruses. The newly developed RT-qPCRs were shown to be more sensitive than a previously described assay in detecting DENV in clinical samples and are suitable for the routine diagnosis.
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Contributor : Philippe Dussart Connect in order to contact the contributor
Submitted on : Sunday, May 9, 2021 - 4:54:21 PM
Last modification on : Tuesday, June 8, 2021 - 2:25:51 PM

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Tey Putita Ou, Chanvannak Yun, Heidi Auerswald, Saraden In, Rithea Leang, et al.. Improved detection of dengue and Zika viruses using multiplex RT-qPCR assays. Journal of Virological Methods, 2020, 282, pp.113862. ⟨10.1016/j.jviromet.2020.113862⟩. ⟨pasteur-03221713⟩



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