Asymmetric GP5+/6+ PCR and hybridization with fluorescence polarization assay of 15 human papillomavirus genotypes in clinical samples

BackgroundThe detection of a broad spectrum of HPV genotypes has not been used widely in cervical cancer screening due to technical difficulties and high costs.ObjectivesTo develop an asymmetric GP5+/6+ PCR assay and hybridization with a fluorescence polarization (FP) assay of 15 HPV genotypes.Study designHPV genes in controls and samples were amplified by an asymmetric GP5+/6+ PCR. Fifteen HPV genotypic probes labeled with fluorophores hybridized with target PCR product to identify the presence of specific HPV genotypes. The HPV genotypes in samples were verified by sequence assay.ResultsThe genotypes determined with the hybridization and FP assay were confirmed by sequence analysis when a monotypic infection was evaluated.ConclusionA simple, economical and specific HPV genotyping assay has been developed that will be adequate for cervical cancer screening programs.