Nakaseomyces glabrata (former name: Candida glabrata) is the second most common cause of vulvovaginal candidiasis (VVC), but its molecular epidemiology and antifungal resistance in China remain poorly understood. This study analysed 204 N. glabrata isolates from VVC patients in Suzhou, Eastern China, using multi-locus sequence typing (MLST) and microsatellite genotyping, alongside antifungal susceptibility testing. A total of 46 sequence types (STs) were identified by MLST, as well as 146 genotypes (GTs) revealed by microsatellite. According to MLST, ST7 was the predominant ST in vaginal N. glabrata isolates, along with a considerably high proportion (32/46, 69.6%) of novel STs. Notably, 27 STs were unique singletons, of which 25 unique STs were newly defined in this investigation. Microsatellite genotyping revealed a similar pattern as MLST with high variability. Population genetic analysis revealed evidence of recombination and ST7 could be the founding population of other related STs. However, there was no significant association between the genotypes and resistance phenotypes. Molecular epidemiology of N. glabrata associated with VVC revealed high genetic variability and the presence of novel genotypes in China. This study highlights the unique genetic profile of vaginal N. glabrata isolates in Suzhou, with the majority of resistant strains belonging to ST7.