Candida tropicalis produces biofilms and other virulence factors like attachment to endothelial and buccal epithelial cell and release of lytic enzymes such as hemolysins, phospholipases, proteinases, and transformation from buds to hyphae (also known as morphogenesis). The aim of this study was to identify Candida tropicalis using API 20 Candida as well as using conventional polymers chain reaction technique for the detection of the genotype and virulence in isolates causing mycotic vaginitis in buffaloes in Wasit Province during the period (May 2021 to October 2021). In this study, one hundred vaginal swab samples were collected from buffalo. The results showed that Candida species were found in 40 out of 100 (40%) of the mycotic vaginal swabs and 30/40 (75%) of the isolated Candida species showed positive results using API 20 Candida test for Candida tropicalis. PCR amplification of the 18S rRNA gene for identification of Candida tropicalis revealed that this gene was detected in 16/30 samples (53.3%). Phospholipase (PLB) gene and hyphal formation (CPH1) gene were both found in 10/16 (62.5%) and 8/16 (50%) of samples respectively. In conclusion, the PCR was a useful tool and excellent alternative method for the early detection and diagnosis of the most frequent Candida species causing infection of genital tract in buffalo where culture was not required.