Type-specific primer-mediated direct sequencing of consensus primer-generated PCR amplicons of human papilloma viruses: a new approach for the simultaneous detection of multiple viral type infections

• Published in: Journal of virological methods Vol. 53; no. 2; pp. 245 - 254
• Main Authors: Rady, Peter L., Arany, Istvan, Hughes, Thomas K., Tyring, Stephen K.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.06.1995
• Subjects: Anus Neoplasms - virology; Base Sequence; Consensus Sequence; Databases, Factual; DNA Primers; DNA, Viral - analysis; DNA, Viral - genetics; HPV; Humans; Molecular Sequence Data; Nasopharyngeal Neoplasms - virology; Oncogene Proteins, Viral - analysis; Oncogene Proteins, Viral - genetics; Papillomaviridae - genetics; Papillomavirus Infections - virology; PCR; Polymerase Chain Reaction - methods; Sequencing; Species Specificity; Tumor Virus Infections - virology; Type-specific primer; Type-specific primer; Sequencing; PCR; HPV
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/0166-0934(95)00029-T

Consensus degenerate primers specific for the L1 and E1 regions were used to amplify anogenital HPV-DNA fragments. The mixture of the viral fragments was then directly sequenced with HPV-6, -11, -16, -18 and -33 type-specific computer-designed oligonucleotides as sequencing primers. The linking of a consensus primer-generated PCR amplification with type-specific primer-mediated direct sequencing and computer data bank analysis provided precise, more objective viral detection in the simultaneous presence of different HPV types. The method was successfully adapted for viral typing in clinical lesions which simultaneously contained different anogenital HPV sequences.