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Positive for HPV 16 Infection ( HPV 16 ) as an Informative Biomarker for the Triage of Women Methylation of Twelve CpGs in Human Papillomavirus Type 16

An accurate biomarker for the follow-up of women positive for human papillomavirus type 16 (HPV16) DNAmay improve the efficiency of cervical cancer prevention. Previously, we analyzed all 113HPV16CpGs in cervical cytology samples and discovered differential methylation at different stages of premalignancy. In the current study, we identified amethylation biomarker consisting of a panel of 12HPV16 CpG sites in the E5, L2, andL1open reading frames, and testedwhether it fulfilled three necessary conditions of a prospective biomarker. A total of 33 cytology samples fromNorth American andWest African womenwith all grades of cervical intraepithelial neoplasia (CIN) and invasive cervical cancer (ICC) were analyzed by using DNA bisulfite sequencing. The results showed (i) a highly significant trend for increasing HPV16 biomarker methylation with increasing histologic severity (P < 0.0001), (ii) 100% sensitivity for ICC over a wide range of methylation cutoff scores; 80% detection of CIN3 at cutoff scores up to 39% methylation, and (iii) substantially lower detection of CIN2, from 0% to 71%, depending on the cutoff score. Our results support the prognostic potential of the HPV16 methylation biomarker for the triage to colposcopy of women with HPV16-positive screening tests and, eventually, for the management of women with HPV16-positive CIN2. Cancer Prev Res; 7(5); 526–33. 2014 AACR.