TYPE-SPECIFIC PRIMER-MEDIATED DIRECT SEQUENCING OF CONSENSUS PRIMER-GENERATED PCR AMPLICONS OF HUMAN PAPILLOMA VIRUSES - A NEW APPROACH FORTHE SIMULTANEOUS DETECTION OF MULTIPLE VIRAL TYPE INFECTIONS

Consensus degenerate primers specific for the L1 and E1 regions were u sed to amplify anogenital HPV-DNA fragments. The mixture of the viral fragments was then directly sequenced with HPV-6, -11, -16, -18 and -3 3 type-specific computer-designed oligonucleotides as sequencing prime rs. The linking of a consensus primer-generated PCR amplification with type-specific primer-mediated direct sequencing and computer data ban k analysis provided precise, more objective viral detection in the sim ultaneous presence of different HPV types. The method was successfully adapted for viral typing in clinical lesions which simultaneously con tained different anogenital HPV sequences.