Concordance analysis of methylation biomarkers detection in self-collected and physician-collected samples in cervical neoplasm

BMC Cancer, May 2015

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. Keywords: Cervical cancer, DNA methylation, Biomarker, Self-collected, Physician-collected, Real-time quantitative methylation-specific polymerase chain reaction (QMSP).

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Concordance analysis of methylation biomarkers detection in self-collected and physician-collected samples in cervical neoplasm

Chang et al. BMC Cancer Concordance analysis of methylation biomarkers detection in self-collected and physician-collected samples in cervical neoplasm Cheng-Chang Chang 1 2 Rui-Lan Huang 0 Yu-Ping Liao 6 Po-Hsuan Su 6 Yaw-Wen Hsu 5 Hui-Chen Wang 6 Chau-Yang Tien 2 Mu-Hsien Yu 2 Ya-Wen Lin 4 Hung-Cheng Lai 0 1 3 0 Department of Obstetrics and Gynecology, Shuang Ho Hospital, Taipei Medical University , Taipei, Taiwan , Republic of China 1 Graduate Institute of Medical Sciences, National Defense Medical Center , Taipei, Taiwan , Republic of China 2 Department of Obstetrics and Gynecology, Tri-Service General Hospital, National Defense Medical Center , Taipei, Taiwan , Republic of China 3 Department of Obstetrics and Gynecology, School of Medicine, College of Medicine, Taipei Medical University , Taipei, Taiwan , Republic of China 4 Department and Graduate Institute of Microbiology and Immunology, National Defense Medical Center , Taipei, Taiwan , Republic of China 5 Graduate Institute of Life Sciences, National Defense Medical Center , Taipei, Taiwan , Republic of China 6 Laboratory of Epigenetics and Cancer Stem Cells, National Defense Medical Center , Taipei, Taiwan , Republic of China 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. Cervical cancer; DNA methylation; Biomarker; Self-collected; Physician-collected; Real-time quantitative methylation-specific polymerase chain reaction (QMSP) - Background Cervical cancer remains one of the main causes of death from cancer among women worldwide [1]. Cytologybased screening has successfully reduced mortality associated with cervical cancer [2]. However, the majority of cases of cervical cancer are still associated with absent or deficient screening. In previous studies, approximately 50 % of cervical cancers were diagnosed in women who were not screened [3, 4]. Complete participation would achieve a greater improvement in screening effectiveness than intensifying screening policies [3]. Therefore, it is important to improve participation rates among women with a history of non-attendance. Epidemiological studies have emphasized that human papillomaviruses (HPVs) are the main etiological factor for cervical cancer and that these viruses are present in almost all cervical cancer tissues [5]. Screening participation rates for cervical cancer can be improved by of (...truncated)


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Cheng-Chang Chang, Rui-Lan Huang, Yu-Ping Liao, Po-Hsuan Su, Yaw-Wen Hsu, Hui-Chen Wang, Chau-Yang Tien, Mu-Hsien Yu, Ya-Wen Lin, Hung-Cheng Lai. Concordance analysis of methylation biomarkers detection in self-collected and physician-collected samples in cervical neoplasm, BMC Cancer, 2015, pp. 418, 15, DOI: 10.1186/s12885-015-1411-x