ABSTRACT: Abnormal vaginal microbiota (AVM), including bacterial vaginosis (BV), is caused by a microbiota imbalance. Nugent scoring is the gold standard for the laboratory diagnosis of BV; however, it is somewhat subjective to interpret, and challenging to distinguish bacteria. Hence, there is a need for improved technologies for the accurate diagnosis of AVM. To this end, next-generation sequencing (NGS) technology has been shown to yield comprehensive information on the pathophysiology of AVM. Hence, to evaluate the relationship between microbiota composition and the pathophysiology of AVM and its clinical significance, we characterized vaginal swab samples from 212 pregnant Korean women using both Nugent scoring and NGS analysis. Of these, the Nugent scoring identified 175 subjects (82.5%; 175/212) with normal flora (NF), 20 (9.4%; 20/212) with intermediate flora (IF), and 17 (8.0%; 17/212) with BV. NGS analysis followed by the characterization of vaginal microbiota composition, as represented by alpha and beta diversity, revealed the relative abundance of specific bacterial taxa at the genus and species level. Moreover, we identified all five predominant community state types (CSTs) along with three smaller CSTs. Analysis of the vaginal microbiota revealed the dominance of one or two Lactobacillus spp. in the NF group. Meanwhile, the IF and BV groups were dominated by the genera Gardnerella, Prevotella, and Atopobium. These two groups also showed higher alpha diversity than the NF group (p < 0.05). Principal coordinate analysis (PCoA) indicated that the NF group was significantly different from the AVM groups (p < 0.05), whereas no significant difference was observed between IF and BV groups (p = 0.25). Lastly, to investigate the characteristics of vaginal microbiota based on taxonomic composition, the IF and BV groups (AVM groups) were reclassified using the unweighted pair group method with arithmetic mean (UPGMA) clustering. Consequently, they were reclassified into BV1 (Lactobacillus iners-dominated), BV2-1 (Bifidobacterium breve-dominated), BV2-2 (Gardnerella vaginalis s1 or s2 and Atopobium vaginae-dominated), and BV3 [mixed population of G. vaginalis, L. iners, and other bacteria (p < 0.05)]. Collectively, these findings could serve to advance the current understanding regarding AVM pathophysiology.