Genotype-Specific HPV E6/E7 mRNA Triage for Risk Stratification in HPV DNA-Positive Women with ASC-US/LSIL: A Population-Based Tromsø Cohort

HPV DNA-positive women with ASC-US/LSIL cytology constitute a heterogeneous triage group. We evaluated genotype-specific HPV E6/E7 mRNA testing with PreTect HPV-Proofer’7 in a population-based cohort from Tromsø and compared the findings descriptively with a previously published cohort from Bodø. This retrospective quality-assurance study included 1006 HPV DNA-positive women with ASC-US/LSIL cytology screened between 2019 and 2024, with linkage to regional pathology records through 31 December 2025. The assay detects E6/E7 mRNA from HPV16, 18, 31, 33, 45, 52, and 58. ASC-US/LSIL accounted for 42.5% of linked HPV DNA-positive women in Tromsø compared with 22.3% in Bodø. Among the mRNA-tested ASC-US/LSIL cohorts, mRNA positivity was similar (40.8% vs. 44.6%). In Tromsø, CIN2+ was recorded in 26.1% of mRNA-positive and 7.9% of mRNA-negative women (RR 3.31, 95% CI 2.41–4.55), whereas CIN3+ was recorded in 3.9% and 1.0%, respectively (RR 3.88, 95% CI 1.53–9.82). For CIN2+, sensitivity was 69.5%, specificity 64.4%, PPV 26.1%, and NPV 92.1%. Among HPV16/18 DNA-positive women, CIN2+ risk was 42.7% in mRNA-positive and 13.4% in mRNA-negative women. Simulated mRNA-guided referral reduced immediate colposcopy referrals by 59.2%. Genotype-specific mRNA testing provided clinically relevant risk stratification, with directionally similar patterns being observed in the Tromsø and Bodø cohorts; however, mRNA-negative women require structured surveillance.