Direct quantification of HIV-1 RNA by capillary electrophoresis with laser-induced fluorescence

J Chromatogr B Biomed Sci Appl. 1997 Sep 12;697(1-2):189-94. doi: 10.1016/s0378-4347(97)00334-4.

Abstract

HIV-1 RNA was quantitated directly by capillary electrophoresis with laser-induced fluorescence (CE-LIF). CE-LIF was used to analyze cellular RNA and various nucleotide complexes. A fluorescently labeled DNA probe (DNA/RNA complex) in conjunction with thiazole orange intercalator was determined to have optimal stability and sensitivity for RNA analysis. Based on this observation, a hybridization method using a HIV-specific fluorescently labeled probe with analysis by CE-LIF was developed. Plasma samples from a HIV-seropositive patient were lysed to obtain RNA, hybridized with the HIV-specific probe and analyzed by CE-LIF. As little as 19 fg (1710 copies per 1 ml of starting plasma) of HIV RNA can be reliably and quantitatively detected. CE-LIF appears to be an efficient and sensitive method to quantitatively analyze RNA from a variety of sources.

MeSH terms

  • Animals
  • Benzothiazoles
  • DNA Probes
  • Electrophoresis, Capillary
  • Fluoresceins
  • Fluorescent Dyes
  • Genes, Viral*
  • HIV-1 / genetics*
  • Humans
  • Intercalating Agents
  • Lasers
  • Nucleotides / analysis
  • Quinolines
  • RNA / analysis
  • RNA / genetics
  • RNA / isolation & purification
  • RNA, Viral / blood*
  • RNA, Viral / genetics
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence
  • Spodoptera / genetics
  • Thiazoles

Substances

  • Benzothiazoles
  • DNA Probes
  • Fluoresceins
  • Fluorescent Dyes
  • Intercalating Agents
  • Nucleotides
  • Quinolines
  • RNA, Viral
  • Thiazoles
  • thiazole orange
  • RNA