Background: High-risk human papilloma virus (hrHPV) is regarded as the main causal factor of cervical precancer and cancer when persistent infection is left untreated. Previous studies have declared that HPV is associated with microecological environment of lower genital tract but haven’t discriminate vaginal microbiota from that of cervix. Objective: To analyze the distinction between vaginal and cervical microbiota in high-risk HPV(+) Chinese women. Methods: One hundred participants were recruited including 20 healthy women with HPV (-), 32 with other hrHPV (+), 38 subjects with HPV16/18 (+) and 10 with cervical carcinoma, demographics of whom were collected and analyzed. Vaginal and cervical microbiota were separately tested through next-generation sequencing technology (NGS) targeting the variable region (V3-V4) of bacterial ribosome 16S rRNA gene. Results: 1. Analysis of demographics demonstrated that hrHPV infected women tend to be accustomed to vaginal douching (p =0.001), show more frequent usage of sanitary pads (p =0.007), have more sex partners (p =0.047), be more sexually active (p =0.025), have more diverse ways of contraception (p =0.001) and history of vaginitis (p =0.002). 2. NGS identified microbial diversity of cervical microbiota was much higher than that of vagina with significantly increased Proteobacteria and decreased Lactobacillus. Variation of cervical microbiota of hrHPV(+) subjects partly similar to vaginal microbiota but had its unique features. Sphingomonas of α-Proteobacteria, almost invisible in the vagina, was more frequent at normal cervix whereas decreased remarkably at hrHPV(+) cervix. Reversely, γ-Proteobacteria showed a significant positive correlation with HPV16/18 and cervical cancer. BV related anaerobes like Gardnerella, Prevotella, Atopobium and Sneathia showed similar changes in both vaginal and cervical microbiota of hrHPV(+) women and did not exhibit cervical specificity. Conclusions: Cervical microbiota has its uniqueness from that of vagina in bacterial communities presenting a higher proportion of Proteobacteria, of which Sphingomonas is potentially predictive of a health guardian of hrHPV while Pseudomonas the opposite.