Objective: To investigate the age-stratified characteristics of high-risk human papillomavirus (hrHPV) infection in women and compare the detection efficacy of three screening protocols in detecting cervical cancer and precancerous lesions across different age groups, to provide a basis for optimizing cervical cancer screening strategies. Methods:? A retrospective analysis was conducted on cervical cancer screening data from 22,540 women aged 35–64 years enrolled in Nanjing’s cervical and breast cancer screening program from 2023 to 2024. Participants were divided into two age groups using the average menopausal age of 50 years as the cut-off: 35–49 years (n=11,539) and 50–64 years (n=11,001). The positive rates of hrHPV 16/18 and the detection rates of cervical intraepithelial neoplasia grade 2 or worse (CIN2+) were compared between the two age groups. Three screening protocols were compared: ThinPrep cytologic test (TCT)-based initial screening, hrHPV-based initial screening, and combined HPV and TCT initial screening (referred to as "combined screening"). The CIN2+ detection rates were compared among the three protocols followed by further age-stratified analysis. The chi-square test was used to compare positive rates and detection rates between groups, and Bonferroni correction (α’=0.016 7) was used for pairwise comparisons if there was a statistically significant difference, with further age-stratified analysis. Logistic regression analysis was applied to adjust for confounding factors and evaluate the differences in screening effectiveness after age stratification. Results:? Age-stratified comparison showed that the positive rate of hrHPV 16/18 was significantly higher in the 50–64 age group than in the 35–49 age group (2.1% vs. 1.6%, χ2=4.672, P=0.031), while there was no statistically significant difference in the CIN2+ detection rate between the two age groups (0.4% vs. 0.5%, χ2=0.210, P=0.647). There was a statistically significant difference in the CIN2+ detection rate among the three screening protocols (χ2=20.999, P<0.001). After Bonferroni correction, pairwise comparisons indicated that both combined screening and HPV initial screening had significantly higher CIN2+ detection rates than TCT initial screening (corrected P< 0.001 for both, P<α’), and there was no statistically significant difference between HPV initial screening and combined screening (corrected P=0.229, P>α’). After age stratification, in the 35–49 age group, the CIN2+ detection rate of combined screening was higher than that of HPV initial screening (0.9% vs. 0.5%, χ2=4.372, P=0.037), whereas in the 50–64 age group, there was no significant difference in the CIN2+ detection rates between the two protocols (0.6% vs. 0.4%, χ2=0.536, P=0.464). The aforementioned age-stratified results were consistent after multivariate logistic regression adjustment. Conclusion:? Combined screening is recommended for women aged 35–49 to improve the detection rate of cervical cancer and precancerous lesions, while HPV initial screening may be prioritized for women aged 50–64. An age-stratified screening strategy can enhance the precision of cervical cancer screening and improve resource utilization efficiency.