The polymerase chain reaction for the detection of HIV-1 genomic RNA in plasma from infected individuals

J Virol Methods. 1991 Feb-Mar;31(2-3):273-83. doi: 10.1016/0166-0934(91)90165-v.

Abstract

HIV (human immunodeficiency virus) viraemia in serum or plasma of HIV-infected individuals was investigated by the polymerase chain reaction assay (PCR) in combination with reverse transcription to detect HIV-1 genomic RNA. Before PCR, plasma or serum was ultracentrifuged, precipitated virions were then treated with a RNase-free DNase, and a cDNA from the HIV-1 genomic RNA was synthesized. Thirty-three fresh plasma and seven sera from either HIV-1 antibody-positive individuals or patients treated with AZT were tested. Plasma from three patients were assayed 3 or 6 months apart. Twelve sera from HIV-1 antibody-negative individuals were used as negative control. PCR was performed with primers in LTR, gag and env regions: 11 of 40 samples were positive with three primer pairs, 16 with two primer pairs and 11 with only one primer pair. PCR on HIV-1 genomic cDNA was positive in 38 out of the 40 plasma or serum samples (95%), regardless of the clinical stage of the infection: HIV-1 was detected in 14 of the 15 untreated subjects and in 24 of the 25 AZT-treated patients. HIV p24 antigen was detected in the serum of 38% of subjects (15 of 40). The results suggest that this method is suitable for the detection of viral particles in plasma or serum from HIV-1-infected individuals irrespective of antiviral treatment.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acquired Immunodeficiency Syndrome / diagnosis
  • Acquired Immunodeficiency Syndrome / drug therapy
  • Cell Line
  • DNA, Viral / blood
  • Deoxyribonucleases
  • Gene Products, env / genetics
  • Gene Products, gag / genetics
  • HIV Long Terminal Repeat
  • HIV-1 / genetics*
  • Humans
  • Polymerase Chain Reaction* / methods
  • Proviruses / genetics
  • RNA, Viral / blood*
  • RNA-Directed DNA Polymerase
  • Sensitivity and Specificity
  • Tumor Cells, Cultured
  • Ultracentrifugation
  • Viremia / drug therapy
  • Zidovudine / therapeutic use

Substances

  • DNA, Viral
  • Gene Products, env
  • Gene Products, gag
  • RNA, Viral
  • Zidovudine
  • RNA-Directed DNA Polymerase
  • Deoxyribonucleases