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Vulvovaginal candidiasis (VVC) alongside antifungal resistance has become a major clinical problem in recent years. A prospective study aimed to evaluate the diversity of yeast strains associated with VVC in the city of Tebessa (northeastern Algeria) and investigate their susceptibility patterns. Over two months, yeasts were isolated using chromogenic media from twenty-nine non-pregnant women with symptomatic VVC. The isolates were characterized using MALDI-TOF MS, and antifungal susceptibility testing was performed for nine antifungal drugs using SensititreTM YeastOneTM YO10. Twenty-nine non-duplicate yeasts were recovered, and the mass spectrometry profiles showed reliable scores, from which four genera and five different species were identified. Candida albicans accounted for 65.5% (n = 19) of the total number of isolates, followed by C. glabrata with 20.7% (n = 6). For the remaining non-albicans Candida (NCA) species, Kluyveromyces marxianus accounted for 6.9% (n = 2), and Pichia kudriavzevii and Saccharomyces cerevisiae had one isolate each. The antifungal susceptibilities showed wild-type MICs of C. albicans to amphotericin B, azoles, and echinocandins. In addition, four C. albicans isolates were resistant to flucytosine. For C. glabrata isolates, a 100% non-WT phenotype was found for both posaconazole and itraconazole. For the very first time, the obtained outcomes provide us with new data about the epidemiology of yeasts causing VVC in Algeria and their antimicrobial susceptibility profiles.