Backgrounds: In this research, an attempt was made to identify Candida isolates collected from women with suspected vulvovaginal candidiasis using single Multiplex PCR reaction as a swift and valid method. Beside, this method was compared with phenotypic methods. Materials & Methods: In this study, 250 vaginal swabs were collected from patients referring to obstetrics and gynecology specialists. In addition to phenotypic methods, multiplex PCR designed by species-specific primers was performed to identify Candida isolates in a single reaction. Descriptive statistics were analyzed by t-test and Chi-square test in SPSS software (Ver. 22) (p<. 05). Findings: According to the results, 92 positive samples were diagnosed using the culture method. Four species were identified by culturing the specimens on CHROM agar. The most common Candida species isolated was C. albicans (54. 3%), followed by C. parapsilosis (28. 2%), C. glabrata (17. 4%), and C. krusei (1. 0%). The most common Candida spp. identified by Multiplex PCR method were C. albicans (50. 0%), C. glabrata (33. 7%), and C. parapsilosis (6. 2%). Also, three mixed infections with C. albicans and C. glabrata as well as C. albicans and C. parapsilosis were identified Conclusion: In comparison to phenotypic methods, considering the cost-effectiveness of PCR methods, the single multiplex PCR reaction was shown to be efficient in epidemiological studies on pathogenic species.