Evaluation of a new non-isotopic sandwich hybridization for the detection of HIV1-specific PCR products

Res Virol. 1995 Jan-Feb;146(1):75-9. doi: 10.1016/0923-2516(96)80592-9.

Abstract

A new non-isotopic sandwich hybridization assay was developed to detect human immunodeficiency virus type 1 (HIV1) provirus amplified by the polymerase chain reaction. The sensitivity and specificity of this new technique using 96-well microplates as the support for the sandwich hybridization procedure and stable enzyme-linked oligomer as the detection probe were compared with those of Southern hybridization using a 32P-labelled oligomer probe. Three laboratories studied 437 peripheral blood mononuclear cell samples from 294 different subjects including both HIV1-seropositive and -seronegative individuals. The non-isotopic assay exhibited a sensitivity of 99.5% and a specificity of 99.1% when compared with the Southern procedure. In addition, the non-isotopic assay gives clear numeric data, is safe when handling, and is especially adapted to large-scale analyses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Blotting, Southern
  • DNA, Viral / analysis*
  • Evaluation Studies as Topic
  • Genes, pol
  • HIV-1 / genetics
  • HIV-1 / isolation & purification*
  • Humans
  • Isotopes
  • Molecular Sequence Data
  • Nucleic Acid Hybridization
  • Polymerase Chain Reaction / methods*
  • Proviruses / genetics
  • Proviruses / isolation & purification
  • Sensitivity and Specificity

Substances

  • DNA, Viral
  • Isotopes