title: Triage of high-risk human papillomavirus-positive women by methylated POU4F3.
abstract: BACKGROUND
  Insufficient specificity of the high-risk human papillomavirus (hrHPV) assay in primary cervical cancer screening results in unnecessary referral. Additional assays to triage hrHPV-positive women are needed to improve molecular cervical cancer screening. DNA methylation is a promising biomarker in cervical cancer. We evaluated the clinical performance of potentially methylated genes as a triage assay for hrHPV-positive women.
  
  
  RESULTS
  We conducted a retrospective hospital-based case-control study in Taiwan. Cervical scrapings were collected before colposcopy for hrHPV testing and quantitative methylation-specific PCR (QMSP) of 16 genes. Five genes, POU4F3, HS3ST2, AJAP1, PAX1, and SOX1, were prioritized for the clinical performance to triage hrHPV-positive women. Two hundred cervical scrapings were randomly classified into a training set (n = 111) and testing set (n = 89). All samples were tested for hrHPV using a Hybrid Capture II (HCII) assay. HrHPV-positive women were subjected to DNA methylation analysis by QMSP. In the training set, the receiver operating characteristic (ROC) curves defined the optimal methylation index (M-index) cutoff values for discriminating CIN3(+) from CIN1/normal, which then were applied to the testing set. Among the five genes, POU4F3 revealed the highest area under the ROC curve (AUC) (0.86; 95 % CI, 0.78-0.95) in detecting CIN3(+). In the testing set, POU4F3 revealed the best clinical performance in triage of hrHPV-positive women with a sensitivity of 74 % and specificity of 89 % for detecting CIN3(+).
  
  
  CONCLUSIONS
  POU4F3 methylation analysis is a potential molecular tool for triage in detecting CIN3(+) in hrHPV-positive women. The combined use of broad-spectrum HPV assay and POU4F3 methylation analysis as a new generation of molecular cervical cancer screening warrants further population-based study.
authors: Molecular Cell Biology, Taiwan International Graduate Program, Academia Sinica, 128 Academia Road, Section 2, Nankang, Taipei, 11529 Taiwan ; Graduate Institute of Life Science, National Defense Medical Center, No.161, Section 6, Min-Chuan East Road, Taipei, 11490 Taiwan.
authors: Par Bahadur
authors: PB
authors: Pun
authors: Department of Obstetrics and Gynecology, School of Medicine, College of Medicine, Taipei Medical University, No. 250, Wuxing Street, Taipei, 11031 Taiwan.
authors: Yu-Ping
authors: YP
authors: Liao
authors: Department of Obstetrics and Gynecology, Shuang Ho Hospital, Taipei Medical University, No. 291, Zhongzheng Rd., Zhonghe District, New Taipei City, 23561 Taiwan.
authors: Po-Hsuan
authors: PH
authors: Su
authors: Department of Obstetrics and Gynecology, School of Medicine, College of Medicine, Taipei Medical University, No. 250, Wuxing Street, Taipei, 11031 Taiwan.
authors: Hui-Chen
authors: HC
authors: Wang
authors: Division of Research and Analysis, Food and Drug Administration, Ministry of Health and Welfare, No. 161-2, Kunyang St, Nangang District, Taipei, 11561 Taiwan.
authors: Yu-Chih
authors: YC
authors: Chen
authors: Graduate Institute of Life Science, National Defense Medical Center, No.161, Section 6, Min-Chuan East Road, Taipei, 11490 Taiwan.
authors: Yaw-Wen
authors: YW
authors: Hsu
authors: Department of Obstetrics and Gynecology, Shuang Ho Hospital, Taipei Medical University, No. 291, Zhongzheng Rd., Zhonghe District, New Taipei City, 23561 Taiwan.
authors: Rui-Lan
authors: RL
authors: Huang
authors: Department of Obstetrics and Gynecology, Tri-Service General Hospital, National Defense Medical, No. 161, Section 6, Min-Chuan East Road, Taipei, 11490 Taiwan.
authors: Cheng-Chang
authors: CC
authors: Chang
authors: Molecular Cell Biology, Taiwan International Graduate Program, Academia Sinica, 128 Academia Road, Section 2, Nankang, Taipei, 11529 Taiwan ; Graduate Institute of Life Science, National Defense Medical Center, No.161, Section 6, Min-Chuan East Road, Taipei, 11490 Taiwan ; Department of Obstetrics and Gynecology, School of Medicine, College of Medicine, Taipei Medical University, No. 250, Wuxing Street, Taipei, 11031 Taiwan ; Department of Obstetrics and Gynecology, Shuang Ho Hospital, Taipei Medical University, No. 291, Zhongzheng Rd., Zhonghe District, New Taipei City, 23561 Taiwan.
authors: Hung-Cheng
authors: HC
authors: Lai
keywords: Biomarker; Cervical cancer screening; Cervical intraepithelial neoplasia (CIN); DNA methylation; HrHPV test; QMSP