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Background: Vulvovaginal candidiasis (VVC) remains one of the most common infections of the female genital tract. This local study was conducted to identify the spectrum of Candida species associated with VVC and recurrent vulvovaginal candidiasis (RVVC) using different phenotypic and genotypic methods and assess their fluconazole susceptibility. Methods: High vaginal swabs were collected from 576 patients with VVC. Swabs were subjected to KOH 10% and cultured on Sabouraud dextrose agar. The species identification of Candida isolates was made using phenotypic methods, including the germ tube test, corn meal agar (CMA), CHROMagar Candida, and PCR-RFLP as the best genotypic methods for confirmation of Candida species in vulvovaginal candidiasis. Fluconazole susceptibility testing was performed using the disk diffusion method. Results: The data were analyzed using the chi-square test. The prevalence of candidiasis was 61%. The socio-demographic profile corresponded to a woman aged 29.14 ± 4.8 years on average, pregnant (77%), diabetic (19%), with hypertension (10%), recurrent (9%), a history of fluconazole use (4%), and under 30 years of age (73%). Vaginal swab cultures yielded Candida growth in 352 cases (61%). Candida albicans was the predominant isolated species (78%), while the most common non-albicans species were C. glabrata (14.7%), C. parapsilosis (4.2%), C. tropicalis (2.8%), and C. krusei (0.56%). Isolates of C. glabrata and C. krusei exhibited the highest rate of Double-Disk Synergy (DDS) resistance among all isolates tested. Conclusion: C. albicans was the most common species. Careful periodical surveillance is needed to identify any changes in the susceptibility patterns to fluconazole with the increased use of this antifungal agent in Sayad Shirazi tertiary care hospital, Iran. [ABSTRACT FROM AUTHOR]