Development of a real-time quantitative RT-PCR for the detection of HIV-2 RNA in plasma
Journal of Virological Methods , Volume 88 - Issue 1 p. 81- 87
An assay is described for the quantification of human immunodeficiency virus type 2 (HIV-2) RNA in EDTA plasma based on RT-PCR using the Taqman real-time PCR detection method. As standard, an electron microscopically counted virus stock of HIV-2 strain NIHZ was used. The lower detection limit is 5 102 HIV-2 RNA copies per ml of EDTA plasma. The assay is linear within the range required (5 102-106 HIV-2 RNA copies/ml of EDTA plasma) with an intra assay variability of 2.5% and an inter-assay variability ranging from 2% at 106 copies to 7.5% at the lower detection limit. Three primer/probe combinations were developed to circumvent false negative samples due to nucleotide variation in the target sequence. Using these primer/probe sets enabled the detection of HIV-2 DNA sequences from all HIV-2 seropositive individuals and two out of five dual human immunodeficiency virus type 1 (HIV-1) and HIV-2 seropositive individuals visiting the University Hospital Rotterdam. (C) 2000 Elsevier Science B.V.
|HIV-2, Plasma viral RNA, Quantitative RT-PCR, Taqman technology|
|Journal of Virological Methods|
|Organisation||Erasmus MC: University Medical Center Rotterdam|
Schutten, M, van den Hoogen, B.G, van der Ende, M.E, Gruters, R.A, Osterhaus, A.D.M.E, & Niesters, H.G.M. (2000). Development of a real-time quantitative RT-PCR for the detection of HIV-2 RNA in plasma. Journal of Virological Methods, 88(1), 81–87. doi:10.1016/S0166-0934(00)00177-4