Objective <p>We evaluated the vaginal microecology and human papillomavirus (HPV) subtype distribution in patients with uterine leiomyoma (UL). The study also assessed the correlation between HPV infection and vaginal microecology dysbiosis and uterine leiomyoma occurrence. We hypothesized that disruption of vaginal microecology and high-risk HPV subtype infections are associated with the occurrence of UL in women of reproductive age.</p> Materials and methods <p>The observational study involved 444 women, divided into two groups: uterine leiomyoma (<i>n</i> = 226) and those without leiomyoma (<i>n</i> = 218). Assessment of vaginal microecological and HPV infection was done on the control and study groups. Spearman correlation was used to analyse differences between the two groups.</p> Results <p>The incidences of uterine leiomyoma in patients aged between 20 and 29 years and 30–39 years were significantly high. UL patients had reduced vaginal cleanliness level (I-II), but a higher number of III-IV degree than in the healthy group. The vulvovaginal candidiasis incidence, vaginal disorders, and infections with HPV16, HPV18, and HPV52 subtypes were also remarkably increased in the patients compared to the controls. The high-risk HPV infection prevalence and vaginal disorders were elevated in the patients compared to the normal group.</p> Conclusions <p>Our findings suggest that HPV infection and vaginal microecological imbalance are associated with the occurrence of uterine leiomyomas. Strategies aimed at maintaining vaginal microecological balance and preventing HPV infection may merit further investigation in relation to uterine leiomyoma management.</p>