Universal ProbeLibrary based real-time PCR assay for detection and confirmation of human papillomavirus genotype 52 infections

摘要

A novel TaqMan real-time PCR (RT-PCR) assay for sensitive and specific detection of HPV 52 infection and confirmation/exclusion of the presence of HPV 52 in clinical specimens positive with a Roche HPV Linear Array cross-reactive probe was developed. Sensitivity of the assay at a 95% detection level was 3.9 DNA copies/reaction and the dynamic range was seven orders of magnitude, discriminating 10-10(7) viral genome equivalents/reaction. Testing 45 HPV-DNA negative samples and 102 HPV-DNA positive samples (42 HPV 52 positive samples and 60 samples containing 30 other HPV genotypes), showed complete agreement with results obtained with GP5+/GP6+ or PGMY09/PGMY0911 PCR-based screening and sequencing. Of the 27 HPV 31/33/58 positive samples cross-reacting with the Linear Array HPV 52 probe, 4 (14.8%) were identified as also containing HPV 52 using RT-PCR. All 16 Linear Array "true HPV 52-positive" samples were confirmed to contain HPV 52 DNA in RT-PCR testing.