Specimen DNA was extracted using the Biomek 3000 workstation (Beckman Coulter, Brea, CA, USA). The β-globin gene was first evaluated by PCR, and only β-globin positive specimens were subjected to HPV evaluation using a pair of SPF1/GP6+ primers14 (link). Sanger sequencing was used for genotyping to evaluate the HPV types, as described previously13 (link)15 (link)16 (link). Samples with ambiguous HPV typing signals were subjected to further cloning and sequencing for multiple infections. The HPV types classified as oncogenic in this study were HPV-16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 6817 (link)18 (link), and all other types including HPV-2, 3, 6, 7, 10, 11, 26, 27, 29, 30, 32, 40, 42, 43, 53, 54, 57, 62, 67, 69, 70, 74, 75, 81, 82, 83, 84, 87, 89, 90, 91, and 94 were classified as non-oncogenic types.
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