Development of an affordable multiplex quantitative RT-PCR assay for early detection and surveillance of Dengue, Chikungunya, and co-infections from clinical samples in resource-limited settings

DENCHIK assay successfully enabled simultaneous amplification of all four DENV serotypes and Chikungunya, from clinical samples. DENCHIK assay detected 7.6% additional Dengue infections and 6.65% fewer Chikungunya infections in clinical samples, demonstrating enhanced diagnostic accuracy. With higher sensitivity and specificity, DENCHIK allows for early detection from day one of symptom onset, improving the estimation of true disease prevalence and mitigating misdiagnosis associated with ELISA-based methods. The integration and surveillance of molecular assays, such as DENCHIK, will enhance epidemiological monitoring of circulating DENV serotypes, CHIKV, and co-infections. These advancements will provide critical insights for public health authorities, enabling them to prioritize treatment, implement effective control measures, and mitigate the transmission of arboviral infections.