Please use this identifier to cite or link to this item:
|Title:||Semi-nested PCR using NS3 primers for the detection and typing of dengue viruses in clinical serum specimens|
NS3 gene primers
|Source:||Seah, C.L.K., Chow, V.T.K., Chan, Y.C. (1995). Semi-nested PCR using NS3 primers for the detection and typing of dengue viruses in clinical serum specimens. Clinical and Diagnostic Virology 4 (2) : 113-120. ScholarBank@NUS Repository. https://doi.org/10.1016/0928-0197(94)00063-Z|
|Abstract:||Background: More rapid, specific and sensitive tests for the laboratory diagnosis of dengue virus infections are needed. Objective: To develop a semi-nested polymerase chain reaction (PCR) assay based on primers within the NS3 gene for the simultaneous detection and typing of dengue viruses in human sera. Study design: A first round of single step reverse transcription-polymerase chain reaction (RT-PCR) was carried out with a pair of consensus primers, followed by a second round of semi-nested amplification using the upstream consensus primer and four type-specific downstream primers. The sensitivity and specificity of the semi-nested PCR assay were determined using plaque- or TCID50-titrated virus-infected tissue culture fluid, and total RNA extracted from C6/36 cells infected with dengue and other flaviviruses, respectively. A retrospective study was performed on acute sera from thirteen patients with dengue (confirmed by virus isolation) employing semi-nested PCR in parallel with virus re-isolation and a single-step RT-PCR method for the typing of dengue viruses in human sera. Results: The semi-nested PCR assay could detect up to 1 pfu of dengue virus, but not other flaviviruses. The semi-nested PCR and single-step RT-PCR assays correctly typed dengue viruses in twelve and five sera, respectively, whereas none of the sera was positive by virus re-isolation. Conclusions: This semi-nested PCR assay is a sensitive and specific tool for the detection and typing of dengue viruses from viremic human sera.|
|Source Title:||Clinical and Diagnostic Virology|
|Appears in Collections:||Staff Publications|
Show full item record
Files in This Item:
There are no files associated with this item.
checked on Jan 22, 2018
WEB OF SCIENCETM
checked on Jan 22, 2018
checked on Jan 20, 2018
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.