Abstract
Zika, yellow fever and dengue viruses are three important flaviviruses that are epidemic and endemic in Central Africa, Middle and South America, Southeast Asia and Pacific Islands. Since these three viral diseases present with similar symptoms and there is a possibility of multiple flaviviruses co-infection, it is important to develop an effective and quick diagnostic tool. Here, we designed and validated a one-step triplex real time reverse transcription polymerase chain reaction (RT-PCR) using TaqMan probes targeting NS5 region for detecting zika virus (ZIKV), dengue virus-4 (DENV-4) and yellow fever virus (YFV). The assay was highly specific for the target viral RNA corresponding to ZIKV, DENV-4 and YFV strains. The sensitivity of the assay was compared to one-step, RT-PCR, it showed a 100-fold higher sensitivity than RT-PCR with ZIKV and DENV and 10-fold higher sensitivity with YFV. In conclusion, the one-step, triplex, TaqMan, real time RT-PCR assay identified and distinguished ZIKV, DENV-4 and YFV by targeting single gene. Key words: Zika, yellow fever, dengue, multiplex real time reverse transcription-polymerase chain reaction (RT-PCR).
Highlights
Zika virus (ZIKV), yellow fever virus (YFV) and dengue virus (DENV) are identified as members of the Flaviviridae family that are transmitted by arthropods
The aim of the current study is to develop a one-step triplex real time reverse transcription polymerase chain reaction (RT-PCR) targeting a different region, the nonstructural protein 5 gene region, for simultaneous detection of ZIKV, DENV and YFV in a single tube
The RNAs of ZIKV, dengue virus-4 (DENV-4) and YFV (Table 2) were amplified by the newly designed triplex TaqMan Real time RT-PCR to evaluate the cross-reactivity of the assay
Summary
Zika virus (ZIKV), yellow fever virus (YFV) and dengue virus (DENV) are identified as members of the Flaviviridae family that are transmitted by arthropods. Aedes aegypti and Aedes albopictus mosquitoes are the common vectors in the transmission of ZIKV, DENV and YFV (Gubler, 1998; Musso et al, 2015). There is currently an overlap in the distribution and vectors transmitting ZIKV, DENV, and YFV.
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