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Title: Rapid, single-step RT-PCR typing of dengue viruses using five NS3 gene primers
Authors: Seah, C.L.K.
Chow, V.T.K. 
Tan, H.C.
Chan, Y.C. 
Keywords: dengue virus
NS3 gene
single-step RT-PCR
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.
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
ISSN: 01660934
DOI: 10.1016/0166-0934(94)00104-O
Appears in Collections:Staff Publications

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