DNA methylation analysis provides a promising triage strategy for cervical intraepithelial neoplasia (CIN) and cancer detection following Human Papillomavirus (HPV) testing on self-collected samples, including urine. This study aimed to develop an entirely molecular cervical screening approach based on HPV and DNA methylation analysis in at-home collected first-void urine from healthy females (n = 69) and a referral population (n = 385; CIN-cancer). CIN3+ detection was analyzed by multivariate logistic regression. Here we show that urinary ASCL1/LHX8 methylation levels increase significantly in relation to disease severity, with AUC-values for CIN3+ of 0.81 (95% CI: 0.74–0.88) and 0.83 (95% CI: 0.74–0.92) in the training (n = 285) and validation cohort (n = 160), respectively. This corresponds to a validated CIN3+ sensitivity of 73.0% (95% CI: 57.0–84.6%) at 81.9% specificity (95% CI: 73.5–88.1%; <CIN2). Urinary HPV testing is more sensitive (83.8%; 95% CI: 68.9–92.3%) although less specific (59.6%; 95% CI: 50.0–68.5%). For triage of HPV positives, ASCL1/LHX8 methylation and HPV16/18 genotyping have a similar CIN3+ sensitivity (75.0%; 95% CI: 62.8–84.2% vs 73.3%; 95% CI: 61.0–82.9%), with lower genotyping specificity. Combining ASCL1/LHX8 methylation with HPV16/18 genotyping yield a 85.0% sensitivity (95% CI: 73.9–91.9%) at 50.5% specificity (95% CI: 40.8–60.1%). The ASCL1/LHX8 methylation test detects nearly all cancers and a majority of CIN3 in first-void urine, supporting the potential of full molecular screening in urine by primary HPV testing and methylation triage. Many women do not participate in cervical cancer screening. Urine self-sampling offers a promising solution to reach these individuals. It allows both HPV-testing (the virus linked to cervical cancer) and a DNA-based follow-up test using the same urine sample, avoiding the need for an extra doctor’s appointment. In this study, we tested this approach using urine samples from 454 women. A clear threshold is set to detect serious cervical issues and cancer. The new DNA-based follow-up test correctly identifies 73% of serious cases, while avoiding false alarms in 82% of healthy individuals. This fully DNA-based urine method makes cervical cancer screening easier and more accessible, especially for those who might otherwise be missed. Van Keer et al. explore an entirely molecular cervical screening approach based on HPV and DNA methylation analysis in at-home collected urine from 454 women. The ASCL1/LHX8 methylation test detects nearly all cancers and a majority of CIN3, supporting the potential of full molecular screening in urine by primary HPV testing and methylation triage.
Keer et al. (Thu,) studied this question.