Triage Performance of FAM19A4/miR124‐2 Methylation: 18‐Month Follow‐Up Results From a Prospective Observational Study Within the Dutch Primary HPV ‐Base

ABSTRACT

DNA methylation analysis of the genes FAM19A4 and miR124‐2 has emerged as a promising triage strategy for high‐risk (hr) human papillomavirus (HPV)‐positive women in cervical screening. This study reports a prospective evaluation of the diagnostic accuracy of FAM19A4/miR124‐2 methylation in a large population‐based primary HPV‐based screening cohort with 18 months of follow‐up. In this prospective observational study, clinician‐collected cervical samples from 3848 consecutive hrHPV‐positive women who participated in the Dutch national primary HPV‐based screening program were tested with the QIAsure Methylation Test, a quantitative methylation‐specific PCR assessing FAM19A4 and miR124‐2 methylation. Through linkage to the Dutch Nationwide Pathology Databank (Palga), 11 cases of cervical carcinoma, 293 cervical intraepithelial neoplasia grade 3 (CIN3), and 29 adenocarcinoma in situ (AIS) were identified during 18 months of follow‐up. Clinical performance for CIN3, AIS, and cancer (CIN3+) detection was assessed based on (I) the threshold of the QIAsure Methylation Test, and (II) thresholds corresponding with clinical specificities of 70% and 80% in this hrHPV‐positive study population. The sensitivity and specificity of FAM19A4/miR124‐2 methylation for CIN3+ detection using the QIAsure Methylation Test threshold (I) were 57.4% (95% CI: 52.0–62.7) and 88.3% (95% CI: 87.2–89.5), respectively. At predefined specificities of 70% and 80%, FAM19A4/miR124‐2 methylation (II) demonstrated a CIN3+ sensitivity of 75.4% (95% CI: 70.7–80.0) and 66.1% (95% CI: 61.0–71.2), respectively. In conclusion, FAM19A4/miR124‐2 methylation analysis showed good triage performance in a primary HPV‐based screening setting. These findings support the use of methylation‐based testing as a triage strategy in primary HPV‐based screening programs, with an appropriate follow‐up policy for hrHPV‐positive women who test methylation‐negative.