High-risk Human Papillomavirus Genotypes in Women Undergoing Cervical Screening in Leshan, China: Real-time and Nested Multiplex PCR

High-risk human papillomavirus (HR-HPV) infection is the principal etiological factor in cervical precancer and cervical cancer. Regional surveillance of HR-HPV prevalence and genotype distribution is important because circulating genotypes may vary according to geography, age structure, screening access, and vaccination coverage. This retrospective cross-sectional study evaluated HR-HPV infection among women undergoing routine cervical cancer screening at Leshan Maternal and Child Health Hospital, Leshan, Sichuan, China, between January 2022 and December 2024. A total of 5,000 eligible women aged 25-60 years were included. Cervical specimens were collected from the transformation zone using a sterile cytobrush and preserved in liquid-based cytology medium. Primary HR-HPV detection was performed using the Roche Cobas 4800 HPV assay, which detects HPV16 and HPV18 individually and detects 12 additional high-risk genotypes as a pooled result. Samples positive for pooled HR-HPV were further analyzed by nested multiplex PCR to identify additional genotypes. Cytology and histopathology results, where available, were reviewed and correlated with HPV detection results. Among 5,000 screened women, 612 were HR-HPV positive, corresponding to an overall prevalence of 12.2%. Single-genotype infections accounted for 65.7% of HPV-positive cases, while multiple-genotype infections accounted for 34.3%. HPV16 was the most frequent genotype, followed by HPV31, HPV56, HPV66, and HPV51. HR-HPV prevalence was highest in women aged 25-30 years and declined with increasing age. Multivariate analysis showed that age below 35 years, multiple sexual partners, smoking, and lack of prior screening were associated with higher odds of HR-HPV positivity. These findings describe the regional epidemiology of HR-HPV infection in a cervical screening cohort from Leshan and support continued genotype surveillance, while conclusions regarding genotype replacement and vaccine impact should be interpreted with caution due to incomplete vaccination history data.