Rapid detection of dengue viral RNA in mosquitoes by nucleic acid-sequence based amplification (NASBA)

RNA amplification by nucleic acid sequence-based amplification (NASBA) was used to detect serotype specific dengue viruses in artificially-infected female Aedes mosquitoes, in comparison with RT-PCR technique. NASBA could detect dengue virus serotype 2 and 4 below 0.1 PFU, which was more sensitive than RT-PCR, but this technique was as sensitive as RT-PCR when detecting dengue virus serotype 1 and 3. Dengue viruses could be detected at the thorax of mosquitoes at 0, 7, and 14 days after inoculation with dengue virus serotype 2. This method should be useful for virological surveillance of dengue infected Aedes mosquitoes, as an early warning system to predict outbreaks of dengue viruses.