Utility of gene methylation analysis, cytological examination, and HPV-16/18 genotyping in triage of high-risk human papilloma virus-positive women

// Yan Tian 1, * , Na-Yi Yuan Wu 2, 3, * , Yu-Ligh Liou 2, 3, 4 , Ching-Tung Yeh 4 , Lanqin Cao 1 , Ya-Nan Kang 1 , Huei-Jen Wang 4 , Yichen Li 4 , Tang-Yuan Chu 5 , Wei Li 6 , Xiang Liu 7 , Yi Zhang 1 , Honghao Zhou 2, 3, * and Yu Zhang 1, * 1 Department of Obstetrics & Gynecology, Xiangya Hospital, Hunan, P. R. China 2 Department of Clinical Pharmacology, Xiangya Hospital, Central South University, Changsha, P. R. China 3 Institute of Clinical Pharmacology, Central South University, Hunan Key Laboratory of Pharmacogenetics, Changsha, P. R. China 4 iStat Biomedical Co. Ltd., Taipei, Taiwan 5 Department of Obstetrics & Gynecology, Buddhist Tzu Chi General Hospital, Hualien, Taiwan 6 Department of Obstetrics and Gynecology, ShengJing Hospital of China Medical University, Liaoning, P. R. China 7 Department of Pharmacy, Xiangtan Central Hospital, Xiangtan, P. R. China * These authors have contributed equally to this work Correspondence to: Yu Zhang, email: cszhangyu@126.com Honghao Zhou, email: hhzhou2003@163.com Keywords: HPV triage, PAX1, ZNF582, HPV16/18 genotyping, cervical cancer Received: June 11, 2016 Accepted: May 11, 2017 Published: July 22, 2017 ABSTRACT In 2015, the American Society for Colposcopy and Cervical Pathology and the Society of Gynecologic Oncology issued interim guidance for the use of a human papillomavirus (HPV) test for primary screening, suggesting triage of women positive for high-risk human papillomavirus (hrHPV) by HPV-16/18 genotyping and cytology for women positive for non-16/18 hrHPV. The design of the present study was based on this interim guidance and analysis of the methylation status of specific candidate genes, which has been proposed as a tool to reduce unnecessary referral following primary HPV screening for cervical cancer. We performed a hospital-based case-control study including 312 hrHPV-positive women. hrHPV genotyping was performed by nested multiplex PCR assay with type-specific primers.Residual cervical cells from liquid-based cytology were used for extraction of genomic DNA for assessment of the methylation status of PAX1 , ZNF582 , SOX1 , and NKX6-1 and HPV genotyping. Combined with HPV-16/18 genotyping, both a dual methylation test for PAX1 / ZNF582 and testing for ZNF582 methylation demonstrated 100% association of methylation with pathology results, indicating carcinoma in situ or squamous cell carcinoma. The sensitivity and specificity of the dual methylation test for PAX1 / ZNF582 as a reflex test for identification of CIN3+ lesions were 78.85% and 73.55% (odds ratio = 10.37, 95% confidence interval = 4.76–22.58), respectively. This strategy could reduce the number of patients referred for colposcopic examination by 31.3% compared with cytology, and thus provide a feasible follow-up solution in regions where colposcopy is not readily available. This strategy could also prevent unnecessary anxiety in women with hrHPV infection.