Sensitive non-radioactive detection of HIV-1: use of nested primers for the amplification of HIV DNA.
This report describes the use of the polymerase chain reaction (PCR) for the non-radioactive detection of HIV-1 proviral genomic sequences in HIV-1 infected cells. We have developed a sensitive assay, using three different sets of nested primers and our results show that this method is superior to standard PCR for the detection of HIV-1 DNA. The assay described features the use of a simple and inexpensive sample preparation technique and a non-radioactive hybridization procedure for confirmation of results. To test the suitability of the assay for clinical purposes, we tested cell samples from 76 anti-HIV-1 positive patients. All were positive for at least one primer set: 88% were positive for all three sets of primers; 9% were positive for two sets of primers and 3% were positive for only one set of primers. It provides a useful approach to the study of HIV-1 infection in patient samples where genomic copies often are present at such low numbers that they are otherwise undetectable.
Teglbjaerg
L L
LL
Department of Infectious Diseases, Hvidovre Hospital, Denmark.
Nielsen
C
C
Hansen
J E
JE
D001483:Base Sequence; D015139:Blotting, Southern; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis*; D005188:False Negative Reactions; D015752:Genes, env; D015750:Genes, gag; D015658:HIV Infections / Q000382:microbiology*; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D011533:Proviruses / Q000302:isolation & purification*
D015342:DNA Probes; D004279:DNA, Viral