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【Abstract】 Objective: By using high-throughput culture and high-throughput sequencing to detect the composition of vaginal microbiota in healthy women and patients with bacterial vaginosis, compare the differences in detection results between the two methods to provide evidence for the role of culturomics in vaginal microecology research and clinical disease diagnosis and treatment. Method: Collect vaginal secretions from 10 healthy women and 10 women with bacterial vaginosis, and use 7 different culture media for high-throughput culture of vaginal secretions. Use Clin-ToF time-of-flight mass spectrometry for identification. Simultaneously, high-throughput sequencing of the 16S rRNA V1V2 variable domain was performed directly on all samples. Result: 1. After culturing 20 vaginal secretion samples using culture omics, 35 bacterial genera were identified, with 98 bacterial species at the species level. A total of 17 Lactobacilli were cultured in all samples, of which 9 were detected in both groups. The top three detected strains in the healthy group were Lactobacillus crispatus, Lactobacillus jensenii, and Lactobacillus iners. And in the 5 samples, both Lactobacillus crispatus and Lactobacillus jensenii were cultured simultaneously. The top three detected in the BV group were Lactobacillus iners, Lactobacillus jensenii, and Lactobacillus psittaci. 2. Using the V1V2 domain of the 16S rRNA gene, high-throughput sequencing was performed on the 20 vaginal microbial samples mentioned above, a total of 18 genera were detected in all samples, of which Lactobacillus genus was detectable in all samples, and the relative abundance of Lactobacillus genus exceeded 50% in 12 samples. 3. Among the 20 vaginal microbiota samples, only 9 bacterial genera were identified simultaneously through culture and high-throughput sequencing. Conclusion: By using high-throughput culturomics and high-throughput sequencing methods to simultaneously detect the vaginal microbiota of all samples, the results showed significant differences. High throughput culture has a higher detection rate of bacterial genera and species, and can cultivate lactobacilli with low or undetectable high-throughput sequencing content. Therefore, the use of high-throughput culture groups will have higher application value in the diagnosis and treatment of bacterial vaginosis and the study of vaginal microbiota.