DNA methylation triage of human papillomavirus-positive atypical squamous cells of undetermined significance in cervical cancer screening
Triage of human papillomavirus (HPV)-positive women with atypical squamous cells of undetermined significance (ASC-US) remains a longstanding clinical challenge in cervical cancer screening, as conventional strategies often lead to excessive colposcopy referrals. We conducted a diagnostic cohort study of 195 HPV-positive women who underwent ASC-US cytology. Multigene DNA methylation analysis was performed on residual cervical exfoliated cells, using histology from colposcopy-directed biopsy and/or endocervical sampling as the reference standard to minimise colposcopy referrals. Our results showed that methylation levels of PAX1, EPB41L3, and FAM19A4 increased progressively from ≤ CIN1 to CIN2, CIN3, and cervical cancer, with significant decreases in ΔCt values from ≤ CIN1 to CIN3 for all three genes ( The host multigene DNA methylation assay demonstrates high specificity and good sensitivity for triaging HPV-positive ASC-US women, with the potential to reduce colposcopy referrals while maintaining robust detection of high-grade lesions, particularly in those aged ≥30 years.