Abstract: Objective To evaluate the value of high-risk human papillomavirus （HR-HPV） DNA detection for cervical precancerous lesions screening in Nanjing. Methods A total of 10 221 cervical samples were performed using multiplex polymerase chain reaction for HR-HPV detection （12 HR types and individual HPV-16／HPV-18 types） and parallel Thinprepcytologic test （TCT）， and meanwhile， 187 colposcopic biopsy were also performed. Results HR-HPV positive rate in 10 221 samples was 10.15％（1037/10 221）， of which HPV 12 types （unspecified） was the most common infective type with the rate of 7.86％. Stratified by age， HR-HPV positive rate in those ＜45 years old was much lower than those≥45 years old with statistic significance（P=0.020）. The detected rate of highgrade cervical intraepithelial neoplasia （CIN） showed tendency to ascend after 45 years old compared with those before 45 years old（P=0.006）. TCT showed that HR-HPV positive rate was 54.22％（199／367）in abnormal cytology group， higher than 8.50％（838／9854）in normal cytology and chronic inflammation group（P=0.000）. Compared with 12 HR types， HPV-16/-18 types showed even more closer with the ascend of cytological lesion（P=00000）. Moreover， a significant correlation was observed between 16 type and the ascending grade of CIN （P=0.003）. Conclusion HR-HPV based screening should be moved forward to 25 years. Moreover， as a high-risk population， women older than 45 years should be closely followed. HPV-16 infection plays an important role in inducing higher grade of cytological lesions which then progress to cervical precancerous lesions. HR-HPV DNA detection can be an alternative screening method to increase the coverage of cervical precancerous lesions screening.