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|Title:||Rapid, single-step RT-PCR typing of dengue viruses using five NS3 gene primers||Authors:||Seah, C.L.K.
|Issue Date:||1995||Citation:||Seah, C.L.K., Chow, V.T.K., Tan, H.C., Chan, Y.C. (1995). Rapid, single-step RT-PCR typing of dengue viruses using five NS3 gene primers. Journal of Virological Methods 51 (2-3) : 193-200. ScholarBank@NUS Repository. https://doi.org/10.1016/0166-0934(94)00104-O||Abstract:||In order to detect and type dengue viruses in serum specimens, four type-specific downstream primers were designed for use with a consensus upstream primer in a reverse transcription and polymerase chain reaction (RT-PCR) assay. RT-PCR using these five primers amplified NS3 gene fragments of diagnostic sizes of 169, 362, 265 and 426 base pairs for dengue virus types 1, 2, 3 and 4, respectively, but not for Japanese encephalitis, Kunjin and yellow fever viruses. The conventional two-step RT-PCR procedure was simplified by combining RT and PCR in a single-step format with a 'hot start'. This RT-PCR protocol was applied successfully to dengue virus-spiked serum and dengue patient serum samples, and could detect as few as one PFU of dengue virus. This assay offers a rapid, specific and sensitive molecular technique for the simultaneous detection and typing of dengue viruses.||Source Title:||Journal of Virological Methods||URI:||http://scholarbank.nus.edu.sg/handle/10635/31305||ISSN:||01660934||DOI:||10.1016/0166-0934(94)00104-O|
|Appears in Collections:||Staff Publications|
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