Dengue emerged as a significant health threat in the endemic regions in recent years. However, inconsistent diagnostic accuracy in sequential dengue infections necessitate improved testing methods to ensure effective management of dengue cases. Here, we evaluated a portable, rapid, and sensitive molecular assay - Reverse Transcriptase Recombinase Polymerase Amplification Assay (RT-RAA) utilizing a mobile suitcase laboratory to detect infections in suspected dengue cases in Bangladesh. 364 suspected dengue patients were enrolled in the study. Dengue cases were confirmed by a positive result from any of the four tests NS1 RDT, IgM RDT, qRT-PCR and RT-RAA assay. IgG RDT was performed to differentiate between primary and secondary dengue infections. Of 364 suspected cases, 320 were confirmed dengue cases, with 55.94% classified as primary and 44.06% as secondary infections. Laboratory results showed comparable positivity rates between RT-RAA(78.8%) and NS1 RDT(77.1%) in primary dengue, followed by qRT-PCR(57.5%) and IgM RDT(12.8%). RT-RAA demonstrated superior positivity rates in secondary dengue(76.6%), surpassing RT-PCR(60.3%), NS1 RDT(27%), and IgM RDT (24.8%). Combining RT-RAA with NS1 RDT detected infections in 89.95% primary and 81.56% secondary dengue. The findings suggest complementing RT-RAA to NS1 RDT could significantly improve dengue detection rate, particularly for secondary infections.
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