Co-amplification of specific sequences of HCV and HIV-1 genomes by using the polymerase chain reaction assay: a potential tool for the simultaneous detection of HCV and HIV-1.
A rapid and simple method using the polymerase chain reaction (PCR) was devised for the co-amplification and simultaneous detection of hepatitis C virus (HCV) and human immunodeficiency virus type 1 (HIV-1) specific sequences in the same serum sample. Genomic RNA was extracted from 13 blood donor sera that were reactive in ELISA for both anti-HCV and anti-HIV-1. The extracted RNA was reverse transcribed into cDNA and amplified using nested primer pairs (SN01 and SN04; SN02 and SN03) based on the HCV prototype sequence of clones 37b and 81, and SK 38/39 for HIV-1 simultaneously. PCR products were analyzed by liquid hybridization or Southern blot hybridization with 32P end-labeled oligonucleotide probes from the regions between the primer pairs, excluding the primer sequences. HCV-RNA was detected in all 13 (100%) samples tested; HIV-RNA was detected in 11 (85%) samples. The ability to co-amplify specific sequences from two different viral genomes in the same reaction mixture offers the possibility of simultaneous detection and diagnosis of more than one viral agent in serum samples of infected individuals.
Nedjar
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Laboratory of Hepatitis, Food and Drug Administration, Bethesda, Maryland 20892.
Biswas
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Hewlett
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D001483:Base Sequence; D015342:DNA Probes; D004277:DNA, Single-Stranded; D015658:HIV Infections / Q000175:diagnosis*; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006526:Hepatitis C / Q000175:diagnosis*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA / Q000097:blood; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic
D015342:DNA Probes; D004277:DNA, Single-Stranded; D012313:RNA