Abstract: OBJECTIVE To investigate the effect of high-risk human papillomavirus (HPV) infections on vaginal Lactobacilli in women. METHODS Sixty-four patients with high-risk HPV infections who were treated in our hospital from Jan. 2014 to Jan. 2016 were selected as observation group, and 64 patients with normal vaginal microecology were selected as control group. Vaginal secretions of the two groups of women were collected. The types of vaginal Lactobacilli and the load of HPV-DNA of the two groups were detected. The number, type and total number of vaginal bacteria were compared between the two groups, and the relationship between the number of Lactobacilli and vaginal cleanliness and HPV-DNA was analyzed. RESULTS The vaginal Lactobacilli count and the ratio of Lactobacilli to total bacterial count in the vagina of observation group were (104.21±70.14) /HP and 0.74±0.02, which were lower than (196.25±75.36) /HP and 0.89±0.07 of control group, and the difference was significant (P<0.05). The Spearman correlation analysis showed a negative correlation between the number of vaginal Lactobacilli and the load of HPV-DNA of the two groups(P<0.05), and a npositive correlation between the vaginal cleanliness and the load of HPV-DNA of the two groups(P<0.05). The vaginal Lactobacilli inert, Lactobacilli and Lactobacilli of the two groups of women had significant differences (P<0.05). CONCLUSION High-risk HPV infection may be closely related to the reduction of vaginal Lactobacilli and the imbalance of bacteria, and increasing the content of Lactobacilli and improving the cleanliness of the vagina can reduce the risk of HPV infection.