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Background and Purpose: Vulvovaginal candidiasis is considered the second most prevalent gynecologic infection among women and one of the main reasons for referring to a gynecologist. During recent decades, probiotic usage has been defined as one of the therapeutic regimens for vaginal candidiasis management, but these findings were controversial. The current study was conducted to determine the effect of fluconazole plus vaginal and oral probiotics supplementation on clinical and mycological improvement of vaginal candidiasis concomitant with antifungal susceptibility of Candida species to fluconazole., Materials and Methods: This double-blind, randomized, placebo-controlled trial was conducted on 76 women with vaginal candidiasis admitted to Naghavi and Imam Reza Gynecology Clinics in Kashan, Central Iran, from July 2017 to March 2020. Patients were diagnosed according to vaginal candidiasis symptoms and positive culture for Candida species. The patients were divided into two groups; one of them received fluconazole plus vaginal and oral probiotics, while the other one received fluconazole with placebo. The clinical and mycological findings were recorded before and after the treatment. In vitro, the fluconazole susceptibility test was determined by the microdilution method according to the Clinical and Laboratory Standards Institute (M27-A3) for the baseline Candida isolates., Conclusion: Based on the findings, 35 days after the intervention, a significant reduction was reported in vaginal candidiasis symptoms in the probiotics supplementation group. Although probiotics supplementation therapy was a better mycological cure, compared to the fluconazole with the placebo group, this difference was not significant (68.4% vs. 46.9%, P =0.184). Exclusion of resistant and susceptible dose-dependent strain in the regression model demonstrated a significant reduction in positive culture probiotics in the supplementation group., Results: Oral and vaginal supplementation with probiotics for 4 weeks played a significant role in the elimination of vaginal candidiasis symptoms. Adjustment of clinical and mycological responses with drug resistance patterns of patients could open a promising horizon for probiotics consumption as a complementary treatment., (Copyright: © Current Medical Mycology.)