To investigate the prevalence, genotype distribution, and associations of HR-HPV infection with cervical lesion grades among Chinese women aged 35-64 years. A cross-sectional analysis was conducted using data from women aged 35-64 years who participated in China's national cervical cancer screening program across 13 provinces in 2021. HR-HPV testing was performed at local laboratories using clinically validated assays approved in China, including pooled detection and genotype-specific platforms following standardized national protocols. Epidemiological characteristics were analyzed in 445,045 women, and genotype distribution was assessed in 113,149 women with available genotyping results. Group differences were assessed using chi-square tests, and factors associated with HR-HPV infection were further explored using multivariable logistic regression. The overall prevalence of HR-HPV infection was 10.42% (95% CI: 10.33-10.51). Significant regional variation was observed, with higher prevalence in Western China (11.48%; 95% CI: 11.30-11.66) and Central China (11.46%; 95% CI: 10.98-11.96), and lower prevalence in Northeastern China (9.12%; 95% CI: 8.89-9.36). HR-HPV prevalence increased with age, peaking in women aged 60-64 years (12.69%; 95% CI: 12.30-13.09). The most prevalent genotypes were HPV52 (2.36%), HPV16 (1.73%), and HPV58 (1.40%). Single infections predominated among HR-HPV positive women (87.91%). HPV16 was the most frequently detected genotype across all cervical lesion grades, while HPV52 (10.35%) and HPV58 (9.69%) accounted for a higher proportion of high-grade lesions than HPV18 (7.27%). HR-HPV infection remains prevalent among Chinese women, and these population-level findings may help inform age- and region-specific cervical cancer screening strategies and provide epidemiological evidence relevant to HPV vaccination planning.