PMID: 11311338Apr 20, 2001Paper

Quantitative assay for group M (subtype A-H) and group O HIV-1 RNA detection in plasma

Journal of Virological Methods
I ErnestJ Remacle

Abstract

A quantitative HIV-1 test is described based on a competitive RT-PCR assay combined with a sandwich hybridization as a detection system. The internal RNA standard (IS) was designed specifically to be competitive during the amplification and during the hybridization step. Sample viral load determination was carried out with one RT-PCR in the presence of 10(3) IS copies. The HIV-1 copy number was calculated by reference to an external standard curve performed on known and increasing amounts of the reference HIV-1 (Ref HIV-1) RNA co-amplified with a constant amount of the IS RNA. The assay had a linear range from 10(1) to 10(6) HIV-1 copies. HIV-1 strains belonging to the different subtypes from group M, but also group O, were all detected. Absolute quantification of purified HIV-1 RNA copies gave identical results as the AMPLICOR HIV-1 Monitor assay. The quantification of patient's samples was evaluated according to different criteria such as dynamic range, sensitivity, efficacy of material recovery, reproducibility and convenience of sample handling. The microplate format of the assay combined with the colorimetric detection provides a convenient tool and fulfills the requirement for routine molecular diagnostic laboratories.

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Citations

Nov 12, 2005·Analytical and Bioanalytical Chemistry·Katie A EdwardsAntje J Baeumner
Sep 23, 2003·Journal of Virological Methods·Marie GueudinFrançois Simon
Jan 31, 2002·AIDS·I D TattJ P Clewley
Mar 21, 2007·Journal of Virological Methods·Makiko HamatakeWataru Sugiura
Jul 6, 2004·Journal of Virological Methods·Bishnupriya BhattacharyaAriberto Fassati
May 18, 2004·Infectious Disease Clinics of North America·Cristian ApetreiStephen M Smith
Jul 11, 2002·Biochemical Pharmacology·Françoise de LonguevilleJosé Remacle

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