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.

Clinical differences among PCR-proven dengue serotype infections.

The objective of this study was to compare the clinical spectra of the dengue serotypes proven by the PCR technique. This retrospective study reviewed the clinical information of dengue-infected patients who were admitted to northeastern provincial hospitals in Thailand from June to September 2002. Dengue infection and viral serotypes were confirmed by polymerase chain reaction (PCR). Paired anti-dengue immunoglobulin G (IgG) and IgM from paired sera were analyzed by enzyme-linked immunosorbent assay (ELISA). Ninety-nine PCR-proven dengue-infected Thai patients were studied. Their ages ranged from 3-30 years. They were infected with DEN1, DEN2, DEN3 and DEN4 in 21, 55, 12, and 12%, respectively. Twenty-two percent had primary and 78% had secondary infections. Dengue fever was the most common presentation for both primary (77.2%) and secondary infections (46.7%). The ratios of dengue fever:dengue hemorrhagic fever (DF:DHF) and non-dengue shock syndrome:dengue shock syndrome (non-DSS:DSS) for DEN2 was the lowest of the dengue serotypes. There was no difference in the duration of fever, percentage of hepatomegaly and bleeding among the serotypes in both DF and DHF. The trends in the white blood cells, lymphocyte and atypical lymphocyte counts in DEN3 were the highest, while those of DEN1 were the lowest of the dengue serotypes.