Replacing an HPV DNA assay with an HPV E6/E7 mRNA full-genotype kit before ART is associated with shorter time to oocyte retrieval without evidence of a lower clinical pregnancy rate: a real-world before–after cohort study

Abstract

Introduction Cervical HPV assessment before assisted reproductive technology (ART) can lead to additional evaluation and delay subsequent treatment. We compared treatment timing and pregnancy outcomes before and after replacement of HPV DNA testing with HPV E6/E7 mRNA full-genotype testing. Methods This single-centre retrospective before–after cohort study included 4,740 women undergoing ART between 1 May 2024 and 31 May 2026. Time to treatment readiness (TTR) was defined as the interval from HPV reporting to subsequent oocyte retrieval within the index ART cycle. The TTR-evaluable cohort included 3,543 women (DNA, n = 3,037; mRNA, n = 506). Between-era differences in TTR were assessed using non-parametric methods and median quantile regression. Clinical pregnancy was evaluated in a nested cohort of 942 women (DNA, n = 732; mRNA, n = 210) using a margin-based safety assessment and logistic regression. Results Median TTR was 30.5 days (IQR 21.0–58.5) in the HPV E6/E7 mRNA era and 63.0 days (IQR 30.0–152.0) in the HPV DNA era ( P < 0.001). The Hodges–Lehmann location-shift estimate was −26.0 days (95% CI −31.0 to −22.0). After adjustment for female age, male age, body mass index (BMI), anti-Müllerian hormone (AMH), and ART cycle status, median TTR was 30.15 days shorter in the HPV E6/E7 mRNA era (95% CI −34.74 to −25.56; P < 0.001). TTR ≥30 days occurred in 53.4% and 75.8% of women, respectively (OR 0.36, 95% CI 0.30–0.44; P < 0.001). Clinical pregnancy occurred in 59.0% of women in the HPV E6/E7 mRNA era and 46.6% in the HPV DNA era. The risk difference was 12.46 percentage points (95% CI 4.89–20.03), and the lower confidence limit exceeded the −5-percentage-point safety margin. The adjusted OR for clinical pregnancy was 1.85 (95% CI 1.33–2.56; P < 0.001). Conclusions HPV E6/E7 mRNA full-genotype testing was associated with a shorter interval to oocyte retrieval than HPV DNA testing. Among pregnancy-evaluable women, the clinical pregnancy rate met the prespecified supportive non-inferiority criterion, with the lower 95% confidence limit for the between-era risk difference exceeding the −5-percentage-point margin.