Although the relationship between vaginal microorganisms and fertility has been well established, only few studies have investigated vaginal microorganisms in women undergoing in vitro fertilization (IVF). Our aim was to study the differences in vaginal microbiota between infertile women with repeated implantation failure (RIF) and those who achieved clinical pregnancy in their first frozen embryo transfer cycle. We compared the vaginal microbiota of patients with a history of RIF (n 5 37) with that of the control group (n 5 43). Following DNA extraction, metagenomic sequencing was employed for the analysis of alpha and beta diversities, distinctions in bacterial species, and the functional annotation of microbial genes. Furthermore, disparities between the two groups were revealed. Alpha diversity analysis revealed that the Shannon index was higher in the RIF group (P < 0.05). There were differences in the beta diversity between groups (P 5 0.16). At the bacterial family level, the relative abundance of Actinomycetaceae (P 5 0.013) and Ruminococcaceae (P 5 0.013) were significantly higher in the RIF group. At the genus level, the abundances of Actinomyces (P 5 0.028) and Subdoligranulum (P 5 0.013) were significantly higher in the RIF group. At the species level, the abundances of Prevotella timonensis (P 5 0.028), Lactobacillus jensenii (P 5 0.049), and Subdoligranulum (P 5 0.013) were significantly higher in the RIF group. Significant differences in family, genus, species, alpha and beta diversity were observed in the vaginal microbiota between groups. Notably, among these findings, the Subdoligranulum genus emerged as the most prominent correlating factor.  Kulcsszavak: RIF, infertility, metagenome, vaginal microbiota, Subdoligranulum