Informative High-Risk HPV Genotyping in Cervical Cancer Screening: Integrated Analysis of Cytology and p16/Ki67 Dual Staining

Background/Objectives: The informative value of integrating high-risk human papillomavirus (HR-HPV) genotyping with cytology and p16/Ki67 dual-stain biomarker results, using limited and two types of extended genotyping assays, has not yet been evaluated. Methods: A total of 32,724 screening test results between 2015 and 2024 were included. Limited HPV genotyping was performed using the Abbott RealTime High Risk HPV assay. Extended genotyping was performed using two assays: the Alinity m HR HPV and BD Onclarity HPV Assay. Trends in age-specific, cytology-specific, and p16/Ki67-specific HR-HPV prevalence and distribution were observed, and differences between limited and extended genotyping were examined. Results: The overall HR-HPV positivity rate was 15.0%. HR-HPV prevalence was 13.9% in the limited genotyping group, 17.8% in in the Onclarity group 1, and 17.2% in the Alinity group 2, with a statistically significant difference in the proportions of positive/negative cases (p < 0.0001). No statistically significant difference was observed between extended genotyping groups (p = 0.706). In the Onclarity group: the highest p16/Ki67 positivity was observed for HPV 33/58 (100.0%) and HPV 31 (58.8%), while the lowest was for HPV 45 (18.2%), HPV 18 (25.0%) and HPV 59/56/66 (28.9%). In the Alinity group: the highest p16/Ki67 positivity was observed for HPV 16 (66.7%) and HPV 31/33/52/58 (58.8%). Conclusions: Based on ten years of HPV-based cervical cancer screening data, this study demonstrates that genotype-specific HR-HPV information obtained through extended genotyping provides clinically relevant risk stratification when interpreted together with cytology and p16/Ki67 dual-stain results. These findings support an integrated screening approach combining molecular HPV testing, cytology, and immunocytochemical biomarkers to improve risk-based triage in cervical cancer screening.