ZNF582 hypermethylation as a highly specific biomarker for triage of high-grade cervical lesions in HR-HPV positive women.

Objective: Persistent infection with high-risk human papillomavirus (HR-HPV) is the primary cause of cervical cancer (CC). DNA methylation is a promising biomarker for cervical cancer screening. This study aimed to validate the clinical efficacy of a cytological DNA methylation-based ZNF 582 methylation (ZNF582m ) test for triage in detecting cervical intraepithelial neoplasia (CIN) grade 3 or higher (CIN3+) among women with HR-HPV infection. Methods: This case-control study examined 199 women who tested positive for HR-HPV from the colposcopy clinic. The cohort included 15 cases of CC, 37 cases of CIN3, 45 cases of CIN2, 49 cases of CIN1, and 53 normal cases. Using cervical pathology results as a reference, the performance of the ZNF582m test for detecting CIN3 + was evaluated and compared with liquid-based cytology (LBC) and P16/Ki67 double stain cytology (DSC). Results: ZNF582m test showed statistically significant differences in pathological results between CIN2 and CIN3 (p < 0.001). The area under the receiver operating characteristic curve (AUC) for distinguishing between <CIN3 and CIN3 + using ZNF582m test was 0.797 (p < 0.001). The ZNF582m test demonstrated higher specificity (88.4, 95% CI, 82.1-93.1%) compared to cytology (≥ atypical squamous cells of undetermined significance; ≥ASC-US+: 28.6, 95% CI: 21.4-36.6%) and P16/Ki67 DSC (43.5, 95% CI, 35.4-52.0%) for detecting CIN3+, establishing it as a superior triage biomarker for women with HR-HPV infection. The Kappa value between the ZNF582m test and pathology (<CIN3 vs. CIN3+) exhibited the highest consistency at 0.589 (0.461-0.716) when compared to other tests, indicating a strong alignment with pathological findings. In the combined test, the specificity of HPV16/18 OR ZNF582m test was found to be the highest among other combined tests, at 63.3% (95% CI = 54.9-71.1%). Conclusion: The ZNF582m test exhibiting superior triage performance compared to P16/Ki67 DSC and LBC (ASC-US+). The ZNF582m test enhances specificity (p < 0.001), thereby facilitating the accurate identification of CIN3 + lesions while reducing unnecessary colposcopies. This is particularly crucial in resource-limited settings, where optimizing the accuracy of triage for HR-HPV positive women is essential.