
Background: The majority of women with Candida vaginitis suffer from uncomplicated vaginitis characterized by sporadic attacks of mild to moderate severity due to C. albicans, and these attacks occur in healthy adult women without any predisposing factors. In contrast, approximately 10% of women suffer from complicated Candida vaginitis, in which attacks either are more severe, occur on a recurrent basis, or are due to non-albicans Candida species. Although several conventional methods are routinely used for determintion; however, Candida vaginitis or vulvovaginitis a challenge for patients and gynecologists. Aims and objectives: Aims of the current study were to find out the prevalence of candidial vulvovaginitis attributed due to Non-Candida albicans Candida (NCAC) species such as Candida glabrata, Candida tropiclis and Candida parapsilosis, and to use a multiplex PCR technique as a rapid identification method for these Non-Candida albicans Candida species isolated from female patients clinically diagnosed with vulvovaginitis. Materials and Methods: In this study vaginal swabs from 160 female patients were used for culture, Api Candida system, VITEK 2 system and multiplex PCR analysis. Multiplex PCR was performed with species-specific primers targeted to the ITS region of rRNA gene of C. glabrata, C. tropicalis and C. parapsilosis. The result of the multiplex PCR was compared with conventional culture, Api Candida system and VITEK 2 system methods. The positive multiplex PCR product was identified by presence of ~ 423 bp, ~357 bp and ~300 bp amplicons of ITS region of rRNA gene for the C. glabrata, C. tropicalis and C. parapsilosis, reaspectively. Results: Conventional methods of candidial culture, Api Candida system and VITEK 2 system, showed that to sum up 100 out of 160 vaginal swabs were detected for Candida species; To sum up 16 out of 100 vaginal swabs were detected for C. glabrata, 10 out of 100 vaginal swabs were detected for C. tropicalis, 6 out of 100 vaginal swabs were detected for C. parapsilosis and 37 out of 100 vaginal swabs were detected for other Candida species. Five of the 60 samples that were negative by conventional methods were positive by multiplex PCR. Statistical analysis revealed that the PCR to have a sensitivity of 95.5 % in the detection of Non-Candida albicans in vulvovaginitis. Conclusion: This multiplex PCR method provides a sensitive, rapid, and reliable alternative to conventional methods to identify Non-Candida albicans Candida species isolated from female patients clinically diagnosed with vulvovaginitis.