DNA-Methylation for Risk-Stratification of Women Without a Fully Visible Transformation Zone at Colposcopy: A Cross-Sectional Study.

Binderup, Karen Omann; Boers, Joachim; Gustafson, Line Winther; Andersen, Berit; Petersen, Lone Kjeld; Bor, Pinar; Gribnau, Joost; Quint, Wim G V et al. · BJOG · 2025

cross_sectional · Level IV

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Abstract

To evaluate the performance of DNA methylation markers for the detection of cervical precancer among screening-positive older women without a fully visible transformation zone at colposcopy (TZ3). Cross-sectional study. Colposcopy clinics, Central Denmark Region, 2019-2021. Women ≥ 45 years with a TZ3 at colposcopy. Participants referred for colposcopy due to an abnormal cervical screening test underwent cervical cytology sampling, biopsies and a large loop excision of the transformation zone (LLETZ). Cytology samples were analysed for DNA methylation markers: FAM19A4/miR124-2, ARID3C/ARL5C and METloc001/METloc002 according to the manufacturer's instructions. LLETZ histology was the reference standard. Sensitivity and specificity of the methylation markers for cervical intraepithelial grade 2 or worse (CIN2+) and cervical intraepithelial grade 3 or worse (CIN3+). Among 89 women (median age: 67.9 years), 31 (34.8%) had CIN2+ and 16 (18.0%) had CIN3+ detected. Sensitivity for CIN2+ was similar across markers, ranging from 74.2% (95% CI: 55.4-88.1) to 77.4% (95% CI: 58.9-90.4), while specificity was lower for FAM19A4/miR124-2 (39.7%; 95% CI: 27.0-53.4) compared to ARID3C/ARL5C (65.5%; 95% CI: 51.9-77.5) and METloc001/METloc002 (82.8%; 95% CI: 70.6-91.4). Sensitivity for CIN3+ was slightly lower for FAM19A4/miR124-2 (75.0%; 95% CI: 47.6-92.7) compared to ARID3C/ARL3C (87.5%; 95% CI: 61.7-98.4) and METloc001/METloc002 (93.8%; 95% CI: 69.8-99.8). Similarly, specificity was lowest for FAM19A4/miR124-2 (35.6%; 95% CI: 24.7-47.7), higher for ARID3C/ARL5C (60.3%; 95% CI: 48.1-71.5) and highest for METloc001/METloc002 (93.8%; 95% CI: 63.9-84.7). Methylation markers, particularly METloc001/METloc002, may be useful for risk-based management of women with a TZ3 at colposcopy. ClinicalTrials.gov identifier: NCT04298957 (https://www. gov/ct2/show/NCT04298957?term=NCT04298957&draw=2&rank=1).