DNA methylation for the triage of HPV-positive clinician-collected cervical samples 67 3 GRAPHICAL ABSTRACT NOVELTY & IMPACT As cervical screening transitions from cytology to primary HPV testing worldwide, effective triage tests are increasingly needed. Here, we report on the performance of host-cell DNA methylation biomarkers ASCL1, LHX8, ST6GALNAC5, GHSR, ZIC1 and SST in a unique, large HPV-positive cohort derived from primary HPV-based screening in the Netherlands. The results show a robust triage performance for CIN3+ as compared to cytology and HPV16/18 genotyping, indicating the strong potential of methylation biomarker-based triage. Routine cervical screening Current study design ROC curve AUC CIN3+ Sensitivity = 76.9% Specificity = 74.5% 1 - Specificity Sensitivity Clinical performance Sample collection DNA extraction Primary HPV test Fluoresence Cycles Positive Negative Threshold Quantitative methylation specific PCR (qMSP) Triage cytology
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