HPV epidemiology and PAX1/JAM3 methylation in cervical cancer screening

Persistent high-risk human papillomavirus (hrHPV) infection drives cervical carcinogenesis, yet accurate biomarkers of high grade cervical intraepithelial neoplasia (CIN2/3) are needed to optimize HPV-based cervical screening. The prevalence of hrHPV types specifically in Changshu was assessed, and the roles of PAX1/JAM3 gene methylation ( PAX1 m /JAM3 m ) in identifying cervical lesions were studied. 52,605 women aged 30–65 years were screened by HPV genotype detection. Furthermore, HPV-positive women were stratified by age and region for subgroup analysis; among them, 173 underwent histopathological assessment, quantitative viral load measurement, and methylation-specific qPCR for PAX1 and JAM3 . The diagnostic performance of HPV 16/18 genotyping, PAX1 m and JAM3 m was compared. We also performed logistic regression models to determine independent predictors of CIN2/3. The overall prevalence of HPV was 6.17% (3245/52605); HPV52 (22.69%, 907/3997), HPV58 (11.56%, 462/3997), and HPV16 (10.13%, 405/3997) were the three most prevalent genotypes. Single genotype infection was detected in 2636 cases (2636/3245, 81.23%), while multiple HPV infections were observed in 609 cases (18.77%). Among multiple HPV infections, 489 cases had two genotypes, 100 had three genotypes, 17 had four genotypes, and 3 had five genotypes. HPV infection prevalence increased with age ( χ² = 101.3, P < 0.0001), peaking at age over 60 years, and differed among sub-districts ( χ² = 61.53, P < 0.0001). Among the 173 follow-up women, including 44 normal lesions, 101 CIN1 lesions, 15 CIN2 lesions, and 13 CIN3 lesions, PAX1 m and JAM3 m levels increased stepwise ( P < 0.0001). Moreover, PAX1 m /JAM3 m had a robust positive association ( R = 0.519, P < 0.0001), but both were independent of viral load. For CIN3, the dual methylation achieved high diagnostic potential, with an area under the curve (AUC) of 0.92. For age-adjusted multivariable analysis, the weighted composite methylation score derived from PAX1 and JAM3 was an independent risk factor for CIN2 + lesions (adjusted OR = 1.965, 95% CI: 1.520–2.541, P < 0.001). The study delineates the HPV prevalence characteristics in the Changshu population. Moreover, it demonstrates that PAX1 m /JAM3 m are viral load-independent biomarkers of CIN2/3 lesions and significantly enhance the diagnostic performance of primary HPV screening.