Belem, L.R.W.; Ibemgbo, S.A.; Gomgnimbou, M.K.; Verma, D.K.; Kaboré, A.; Kumar, A.; Sangaré, I.; Sunil, S. Development of Multiplex Molecular Assays for Simultaneous Detection of Dengue Serotypes and Chikungunya Virus for Arbovirus Surveillance. Curr. Issues Mol. Biol.2024, 46, 2093-2104.
Belem, L.R.W.; Ibemgbo, S.A.; Gomgnimbou, M.K.; Verma, D.K.; Kaboré, A.; Kumar, A.; Sangaré, I.; Sunil, S. Development of Multiplex Molecular Assays for Simultaneous Detection of Dengue Serotypes and Chikungunya Virus for Arbovirus Surveillance. Curr. Issues Mol. Biol. 2024, 46, 2093-2104.
Belem, L.R.W.; Ibemgbo, S.A.; Gomgnimbou, M.K.; Verma, D.K.; Kaboré, A.; Kumar, A.; Sangaré, I.; Sunil, S. Development of Multiplex Molecular Assays for Simultaneous Detection of Dengue Serotypes and Chikungunya Virus for Arbovirus Surveillance. Curr. Issues Mol. Biol.2024, 46, 2093-2104.
Belem, L.R.W.; Ibemgbo, S.A.; Gomgnimbou, M.K.; Verma, D.K.; Kaboré, A.; Kumar, A.; Sangaré, I.; Sunil, S. Development of Multiplex Molecular Assays for Simultaneous Detection of Dengue Serotypes and Chikungunya Virus for Arbovirus Surveillance. Curr. Issues Mol. Biol. 2024, 46, 2093-2104.
Abstract
The major arboviruses belong mainly to the families Bunyaviridae, Togaviridae, and Flaviviridae amongst which chikungunya virus and dengue virus have emerged as a global public health problem. The main objective of the study was to develop a specific, sensitive, and cost-effective molecular multiplex RT-PCR and RT-qPCR assay for the rapid and simultaneous detection of CHIKV and the four serotypes of DENV for arbovirus surveillance. Specific primers of all virus were designed, one step multiplex RT-PCR (mRT-PCR) and RT-qPCR (mRT-qPCR) was developed using reference strains of CHIKV and DENV serotypes. The specificity of the test for all the viruses was confirmed by sequencing. The standard curves showed a high correlation coefficient, R2= 0.99, for DENV-2 and DENV-3; R2= 0.98, for DENV-4 and CHIKV; R2= 0.93, for DENV-1. The limits of detection were calculated to be 10-2 copies/µL for all viruses. The specificity and sensitivity of the newly developed mRT-PCR and mRT-qPCR were validated using positive serum samples collected in India and Burkina Faso. The sensitivity of mRT-PCR and mRT-qPCR are 91%, and 100% respectively. The specificity of both assays was 100% specific. mRT-PCR and mRT-qPCR assays are low cost and a combination of both will be a useful tool for arbovirus surveillance.
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