title: Concordance analysis of methylation biomarkers detection in self-collected and physician-collected samples in cervical neoplasm.
abstract: BACKGROUND
  Non-attendance at gynecological clinics is a major limitation of cervical cancer screening and self-collection of samples may improve this situation. Although HPV testing of self-collected vaginal samples is acceptable, the specificity is inadequate. The current focus is increasing self-collection of vaginal samples to minimize clinic visits. In this study, we analyzed the concordance and clinical performance of DNA methylation biomarker (PAX1, SOX1, and ZNF582) detection in self-collected vaginal samples and physician-collected cervical samples for the identification of cervical neoplasm.
  
  
  METHODS
  We enrolled 136 cases with paired methylation data identified from abnormal Pap smears (n = 126) and normal controls (n = 10) regardless of HPV status at gynecological clinics. The study group comprised 37 cervical intraepithelial neoplasm I (CIN1), 23 cervical intraepithelial neoplasm II (CIN2), 16 cervical intraepithelial neoplasm III (CIN3), 30 carcinoma in situ (CIS), 13 squamous cell carcinomas (SCCs) and seven adenocarcinomas (ACs)/adenosquamous carcinomas (ASCs). PAX1, SOX1 and ZNF582 methylation in study samples was assessed by real-time quantitative methylation-specific polymerase chain reaction analysis. We generated methylation index cutoff values for the detection of CIN3+ in physician-collected cervical samples for analysis of the self-collected group. Concordance between the physician-collected and self-collected groups was evaluated by Cohen's Kappa. Sensitivity, specificity and area under curve (AUC) were calculated for detection of CIN3+ lesions. Finally, we produced an optimal cutoff value with the best sensitivity from the self-collected groups.
  
  
  RESULTS
  We generated a methylation index cutoff value from physician-collected samples for detection of CIN3+. There were no significant differences in sensitivity, specificity of PAX1, SOX1 and ZNF582 between the self-collected and physician-collected groups. The methylation status of all three genes in the normal control samples, and the CIN 1, CIN2, CIN3, CIS, ACs/ASCs and SCC samples showed reasonable to good concordance between the two groups (κ = 0.443, 0.427, and 0.609 for PAX1, SOX1, and ZNF582, respectively). In determining the optimal cutoff values from the self-collected group, ZNF582 showed the highest sensitivity (0.77; 95%CI, 0.65-0.87) using a cutoff value of 0.0204.
  
  
  CONCLUSIONS
  Methylation biomarker analysis of the three genes for detection of CIN3+ lesions shows reasonable to good concordance between the self-collected and physician-collected samples. Therefore, self-collection of samples could be adopted to decrease non-attendance and improve cervical screening.
authors.affiliation: Department of Obstetrics and Gynecology, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan, Republic of China. sundoor66@gmail.com.|Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan, Republic of China. sundoor66@gmail.com.
authors.forename: Cheng-Chang
authors.initials: CC
authors.lastname: Chang
authors.affiliation: Department of Obstetrics and Gynecology, Shuang Ho Hospital, Taipei Medical University, Taipei, Taiwan, Republic of China. gyntsgh@gmail.com.
authors.forename: Rui-Lan
authors.initials: RL
authors.lastname: Huang
authors.affiliation: Laboratory of Epigenetics and Cancer Stem Cells, National Defense Medical Center, Taipei, Taiwan, Republic of China. lypingg@gmail.com.
authors.forename: Yu-Ping
authors.initials: YP
authors.lastname: Liao
authors.affiliation: Laboratory of Epigenetics and Cancer Stem Cells, National Defense Medical Center, Taipei, Taiwan, Republic of China. pohsuan@ms8.hinet.net.
authors.forename: Po-Hsuan
authors.initials: PH
authors.lastname: Su
authors.affiliation: Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan, Republic of China. Fri13wen@gmail.com.
authors.forename: Yaw-Wen
authors.initials: YW
authors.lastname: Hsu
authors.affiliation: Laboratory of Epigenetics and Cancer Stem Cells, National Defense Medical Center, Taipei, Taiwan, Republic of China. egg-0420@yahoo.com.tw.
authors.forename: Hui-Chen
authors.initials: HC
authors.lastname: Wang
authors.affiliation: Department of Obstetrics and Gynecology, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan, Republic of China. TING711212@hotmail.com.
authors.forename: Chau-Yang
authors.initials: CY
authors.lastname: Tien
authors.affiliation: Department of Obstetrics and Gynecology, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan, Republic of China. hsienhui@ms15.hinet.net.
authors.forename: Mu-Hsien
authors.initials: MH
authors.lastname: Yu
authors.affiliation: Department and Graduate Institute of Microbiology and Immunology, National Defense Medical Center, Taipei, Taiwan, Republic of China. ndmc.yawen@msa.hinet.net.
authors.forename: Ya-Wen
authors.initials: YW
authors.lastname: Lin
authors.affiliation: Graduate Institute of Medical Sciences, National Defense Medical Center, Taipei, Taiwan, Republic of China. hclai30656@gmail.com.|Department of Obstetrics and Gynecology, Shuang Ho Hospital, Taipei Medical University, Taipei, Taiwan, Republic of China. hclai30656@gmail.com.|Department of Obstetrics and Gynecology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan, Republic of China. hclai30656@gmail.com.
authors.forename: Hung-Cheng
authors.initials: HC
authors.lastname: Lai
mesh_terms: D000328:Adult; D014408:Biomarkers, Tumor; D002278:Carcinoma in Situ / Q000235:genetics / Q000473:pathology; D019175:DNA Methylation; D055088:Early Detection of Cancer; D044127:Epigenesis, Genetic; D057890:Epigenomics; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D008875:Middle Aged; D009367:Neoplasm Staging; D016133:Polymerase Chain Reaction; D012372:ROC Curve; D012016:Reference Values; D015203:Reproducibility of Results; D002583:Uterine Cervical Neoplasms / Q000235:genetics* / Q000473:pathology; D014626:Vaginal Smears; D002578:Uterine Cervical Dysplasia / Q000235:genetics / Q000473:pathology
chemical_list: D014408:Biomarkers, Tumor