High-risk human papillomavirus (hrHPV) infection is an essential factor in the development of cervical cancer, making timely treatment of this infection important. The number of individuals with persistent hrHPV in the gynecological outpatient setting remains high, and no effective hrHPV drug has been brought to market. Previously, Rebacin, an anti-HPV gel dressing, was shown to be a safe and effective drug for clearing persistent hrHPV infections; however, larger real-world studies examining safety and accounting for HPV subtype and patients age are needed. This prospective multicenter cohort study examined the safety and efficacy of Rebacin for the treatment of persistent hrHPV infections and analyzed impact of hrHPV subtypes, single/multiple-type infections, and patient age. Patients were recruited from 239 hospitals across 21 provinces of China. Premenopausal adult women with persistent hrHPV for greater than12 months without CIN 2 or above were included. Based on the treating physician's recommendation and the patient's preference, eligible patients were allocated either to the Rebacin cohort or a control cohort receiving no treatment. Those in theRebacin cohort received intravaginal Rebacin every other day for 3 months (1 course), except during the menstrual period. Testing for HPV infection was done with 3 routine methods at baseline and 28-35 days following withdrawal of Rebacin, including HPV genotyping, HPV HC II load, or HPV E6/E7 mRNA detection. A total of 3501 patients met inclusion criteria between July 2018 and November 2022. Significantly more participants in the Rebacin cohort had their infection cleared after 1 course of treatment than in an equal amount of time among the control cohort (60% [1972/3252] vs 20.08% [50/249];P< 0.001). In the Rebacin cohort, the clearance rates of hrHPV subtypes 16,18, 58, and 52 were found to be 70.62%, 69.23%, 62.71%, and 59.57%, respectively. Clearance rates were significantly affected by the number of hrHPV subtypes present, where patients with a single, double, triple infections had clearance rates of65.34%, 53.31%, and 38.60%, respectively (P< 0.001). Patients younger than 40 had a significantly greater clearance ratethan those over 40 (65.14% [1065/1635] vs 55.08% [797/1447];P< 0.001). No serious adverse events were reported. The results of this study suggest that after a single 3-month course of Rebacin, a significantly higher clearance rate of hrHPV infections was observed compared with a control group, and factors such as hrHPV subtype, patient age >40, and presence of multiple infection subtypes adversely affected clearance rates.