Cavuslu S, Starkey W G, Kaye J N, Biswas C, Mant C, Kell B, Rice P, Best J M, Cason J
Richard Dimbleby Laboratory of Cancer Virology, Department of Virology, Rayne Institute, United Medical School, St Thomas' Hospital, London, UK.
J Virol Methods. 1996 Apr 26;58(1-2):59-69. doi: 10.1016/0166-0934(95)01988-x.
The development of a nested polymerase chain reaction (PCR) assay to detect low concentrations of human papillomavirus type-16 (HPV-16) DNA for epidemiological studies is described. The PCR utilises primers located in the E5 open reading frame, has an analytical sensitivity of 4 HPV-16 genomes and does not produce amplicons from other common genital HPVs (types-6, -11, -18, -31 and 33). This assay was carried out in 96-well plates utilising internal primers labelled with dinitrophenol (DNP) and biotin so that amplicons can be captured onto streptavidincoated plates and detected using an alkaline phosphatase-labelled monoclonal antibody to DNP. The assay was effective for detecting HPV-16 DNA in plasmids, cell-lines and, both freshly collected or archival (formalin-fixed/paraffin embedded) clinical specimens. This system is therefore suitable for epidemiological studies to identify individuals infected with HPV-16 DNA in episomal form who may be at increased risk of developing anogenital carcinomas.
