title: Comparison of HPV genotyping and methylated ZNF582 as triage for women with equivocal liquid-based cytology results.
abstract: BACKGROUND
  The interpretation of equivocal Papanicolaou (Pap) smear results remains challenging, even with the addition of the high-risk human papillomavirus test (HPV-HR). Recently, methylated zinc finger protein 582 (ZNF582) (ZNF582 (m) ) was reported to be highly associated with cervical cancer. In this study, we compared the performance of ZNF582 (m) detection and HPV-HR genotyping in the triage of cervical atypical squamous cell of undetermined significance (ASC-US) and atypical squamous cell - cannot exclude a high-grade lesion (ASC-H).
  
  
  METHODS
  Two hundred and forty-two subjects with equivocal papanicolaou smear (Pap smear) results were recruited in this hospital-based and case-controlled study. The residual cervical cells in liquid-based cytological test (LBC) containers were used for genomic DNA extraction and then for ZNF582 (m) and HPV-HR detection. The level of ZNF582 (m) was quantified by real-time methylation-specific PCR after bisulfite conversion. The HPV-HR test was performed by using a nested multiplex PCR (NMPCR) assay that combines degenerate E6/E7 consensus primers and HPV type-specific primers.
  
  
  RESULTS
  Significant associations were observed between ZNF582 (m) and the risk of cervical intraepithelial neoplasia grade 3 or higher (CIN3+; odds ratio = 15.52, 95% confidence interval (CI): 7.73 to 31.18). The sensitivity and specificity of ZNF582 (m) for women with CIN3+ were 82.43% and 76.79%, respectively. High sensitivity (99.33%) but low specificity (38.76%) was observed for HPV-HR. When combining both positive results of ZNF582 (m) and HPV-HR, the sensitivity and specificity were 82.43% and 81.55%, respectively. The sensitivity and specificity of ZNF582 (m) or HPV-16/18 were 89.19% and 70.24%, respectively. However, the sensitivity and specificity of ZNF582 (m) combined with HPV-16/18 (both ZNF582 (m) and HPV-16/18 positive results) were 59.46% and 94.64%, respectively.
  
  
  CONCLUSIONS
  ZNF582 (m) provides a promising triage tool for women with ASC. To effectively manage ASC patients, a new strategy co-testing for ZNF582 (m) and HPV-16/18 genotyping was proposed. This strategy could reduce the number of patients referred for colposcopic examination and thus provide a feasible follow-up solution in the regions where colposcopy is not readily available. This strategy could also prevent women from experiencing unnecessary anxiety caused by HPV-HR.
authors.affiliation: Department of Clinical Pharmacology, Xiangya Hospital, Central South University, No. 110 Xiangya Road, Changsha, Hunan 410008 People's Republic of China ; Institute of Clinical Pharmacology, Central South University, Hunan Key Laboratory of Pharmacogenetics, No. 110 Xiangya Road, Changsha, Hunan 410078 People's Republic of China ; iStat Biomedical Co. Ltd., 18F, No. 96, Sec.1, Xintai 5th Road, Xizhi Dist., Taipei, 22102 Taiwan.
authors.forename: Yu-Ligh
authors.initials: YL
authors.lastname: Liou
authors.affiliation: Department of Obstetrics and Gynecology, Xiangya Hospital, Central South University, No. 87 Xiangya Road, Changsha, Hunan 410008 People's Republic of China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Clinical Pharmacology, Xiangya Hospital, Central South University, No. 110 Xiangya Road, Changsha, Hunan 410008 People's Republic of China ; Institute of Clinical Pharmacology, Central South University, Hunan Key Laboratory of Pharmacogenetics, No. 110 Xiangya Road, Changsha, Hunan 410078 People's Republic of China.
authors.forename: Yingzi
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Obstetrics and Gynecology, Xiangya Hospital, Central South University, No. 87 Xiangya Road, Changsha, Hunan 410008 People's Republic of China.
authors.forename: Lanqin
authors.initials: L
authors.lastname: Cao
authors.affiliation: Department of Clinical Pharmacology, Xiangya Hospital, Central South University, No. 110 Xiangya Road, Changsha, Hunan 410008 People's Republic of China.
authors.forename: Chong-Zhen
authors.initials: CZ
authors.lastname: Qin
authors.affiliation: Department of Clinical Pharmacology, Xiangya Hospital, Central South University, No. 110 Xiangya Road, Changsha, Hunan 410008 People's Republic of China.
authors.forename: Tao-Lan
authors.initials: TL
authors.lastname: Zhang
authors.affiliation: iStat Biomedical Co. Ltd., 18F, No. 96, Sec.1, Xintai 5th Road, Xizhi Dist., Taipei, 22102 Taiwan.
authors.forename: Chi-Feng
authors.initials: CF
authors.lastname: Chang
authors.affiliation: iStat Biomedical Co. Ltd., 18F, No. 96, Sec.1, Xintai 5th Road, Xizhi Dist., Taipei, 22102 Taiwan.
authors.forename: Huei-Jen
authors.initials: HJ
authors.lastname: Wang
authors.affiliation: Department of Education and Research, Taipei City Hospital, No.10, Sec. 4, Ren'ai Rd., Da'an Dist., Taipei, 10629 Taiwan.
authors.forename: Shu-Yi
authors.initials: SY
authors.lastname: Lin
authors.affiliation: Department of Obstetrics and Gynecology, Buddhist Tzu Chi General Hospital, No.707, Sec. 3, Zhongyang Rd., Hualien, 97002 Taiwan ; Institure of Medical Science, Tzu Chi University, No.707, Sec. 3, Zhongyang Rd., Hualien, 97002 Taiwan ; Center for Cervical Cancer Prevention, Department of Research Buddhist Tzu Chi General Hospital, No.707, Sec. 3, Zhongyang Rd., Hualien, 97002 Taiwan.
authors.forename: Tang-Yuan
authors.initials: TY
authors.lastname: Chu
authors.affiliation: Department of Obstetrics and Gynecology, Xiangya Hospital, Central South University, No. 87 Xiangya Road, Changsha, Hunan 410008 People's Republic of China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Clinical Pharmacology, Xiangya Hospital, Central South University, No. 110 Xiangya Road, Changsha, Hunan 410008 People's Republic of China ; Institute of Clinical Pharmacology, Central South University, Hunan Key Laboratory of Pharmacogenetics, No. 110 Xiangya Road, Changsha, Hunan 410078 People's Republic of China.
authors.forename: Hong-Hao
authors.initials: HH
authors.lastname: Zhou
keywords: Biomarker; Cervical cancer; DNA methylation; HPV-16/18; HPV-HR; ZNF582