The polymerase chain reaction for the detection of HIV-1 genomic RNA in plasma from infected individuals.
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.
Ottmann
M
M
Laboratoire de virologie, Faculté de médecine-Centre Hospitalier Universitaire, Grenoble, France.
Innocenti
P
P
Thenadey
M
M
Micoud
M
M
Pelloquin
F
F
Seigneurin
J M
JM
D000163:Acquired Immunodeficiency Syndrome / Q000175:diagnosis / Q000188:drug therapy; D002460:Cell Line; D004279:DNA, Viral / Q000097:blood; D003851:Deoxyribonucleases; D015686:Gene Products, env / Q000235:genetics; D015683:Gene Products, gag / Q000235:genetics; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D011533:Proviruses / Q000235:genetics; D012367:RNA, Viral / Q000097:blood*; D012194:RNA-Directed DNA Polymerase; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured; D014461:Ultracentrifugation; D014766:Viremia / Q000188:drug therapy; D015215:Zidovudine / Q000627:therapeutic use
D004279:DNA, Viral; D015686:Gene Products, env; D015683:Gene Products, gag; D012367:RNA, Viral; D015215:Zidovudine; D012194:RNA-Directed DNA Polymerase; D003851:Deoxyribonucleases