Abstract: OBJECTIVE To investigate the infuential factors of vaginal mciroecological testing on the outcome of human papillomavirus（HPV） infection in patients with high-grade squamous intraepithelial lesions（HSIL）. METHODS A total of 132 patients with HISL-HPV infection diagnosed by pathology combined with HPV-DNA testing in the first affiliated hospital of Jinzhou Medical University from Dec. 2013 to Dec. 2018 were selected as the research subjects, and 120 healthy women undergoing physical examination in the hospital were selected as the control group. After 12 months of diagnosis, the vaginal microecological detection method was used to evaluate the various indicators of vaginal secretions in the study group. The relationship between HSIL-HPV outcome and vaginal microecology and its influencing factors were analyzed. RESULTS Totally 132 patients with HSIL-HPV were re-examined, 104 HSIL-HPV patients became negative, and 28 patients were continuously infected with HPV. The proportion of normal vaginal flora in the vaginal transformation group was significantly higher than that in the HPV persistent infection group, but significantly lower than that in the control group（P＜0.05）. The normal density and diversity of the bacteria in the HPV negative transition group were 93.27%（97/104） and 94.23%（98/104）, respectively, significantly higher than those in the HPV persistent infection group（P＜0.05）. Treatment method and the initial viral load were the influential factors for the prognosis of HSIL-HPV infection in female genital tract. CONCLUSION Non-surgical treatment and high viral load of HSIL patients with HPV have higher persistent infection rate, and the vaginal microecological evaluation system is of significance for changes in intravaginal infection of HPV patients with high-grade cervical lesions.