Development of a real-time quantitative RT-PCR for the detection of HIV-2 RNA in plasma
| Autor: | Hubert G. M. Niesters, B G van den Hoogen, Martin Schutten, M.E. van der Ende, A.D.M.E. Osterhaus, Rob A. Gruters |
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| Přispěvatelé: | Virology, Erasmus MC other |
| Rok vydání: | 2000 |
| Předmět: |
Detection limit
Reverse Transcriptase Polymerase Chain Reaction RNA HIV Infections Viral Load Biology Virology Molecular biology Reverse transcriptase Virus chemistry.chemical_compound Real-time polymerase chain reaction chemistry SDG 3 - Good Health and Well-being HIV-2 HIV-1 TaqMan Humans RNA Viral Taq Polymerase Viral load Taq polymerase |
| Zdroj: | Journal of Virological Methods, 88, 81-87. Elsevier |
| ISSN: | 0166-0934 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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