Quantitation of HIV-1 DNA with a sensitive TaqMan assay that has broad subtype specificity

J Virol Methods. 2013 Jan;187(1):94-102. doi: 10.1016/j.jviromet.2012.09.019. Epub 2012 Oct 8.

Abstract

The increasing diversity of HIV-1 isolates makes virus quantitation challenging, especially when diverse isolates co-circulate in a geographical area. Measuring the HIV-1 DNA levels in cells has become a valuable practical tool for fundamental and clinical research. A quantitative HIV-1 DNA assay was developed based on TaqMan(®) technology. Primers that target the highly conserved LTR region were designed to detect a broad array of HIV-1 variants, including viral isolates from many subtypes, with high sensitivity. Introduction of a pre-amplification step prior to the TaqMan(®) reaction allowed the specific amplification of fully reverse transcribed viral DNA. Execution of the pre-amplification step with a second primer set enables for the exclusive quantitation of the 2-LTR circular HIV-1 DNA form.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • DNA Primers
  • DNA, Viral / analysis*
  • DNA, Viral / genetics
  • Genetic Variation
  • HIV Long Terminal Repeat
  • HIV-1 / classification
  • HIV-1 / genetics*
  • HIV-1 / isolation & purification*
  • Humans
  • In Situ Hybridization, Fluorescence
  • Polymerase Chain Reaction*
  • RNA, Viral
  • Sensitivity and Specificity
  • T-Lymphocytes / virology
  • Taq Polymerase

Substances

  • DNA Primers
  • DNA, Viral
  • RNA, Viral
  • Taq Polymerase