Trichomoniasis is a sexually transmitted disease caused by a mobile flagellate pathogenic protist protozoan of the urogenital tract, Trichomonas vaginalis. No data is available in Côte d'Ivoire about its diagnosis by molecular methods. This study aims to identify T. vaginalis by polymerase chain reaction (PCR) among women at Institut Pasteur of Côte d'Ivoire. Vaginal swabs were obtained from each woman from July to October, 2013. For T. vaginalis detection, Giemsa stain for microscopic examination and real-time PCR were performed. The PCR targeted 67 bp region of a repeated sequence of the T. vaginalis genome. A positive T. vaginalis result was defined as a cycle threshold (Ct) less than 36. The results show that of the 194 specimens tested by both Giemsa stain method and Real-time PCR, 2 were positive to Giemsa stain (1.03%) and 7 were positive in PCR assay (3.61%). Women who had multiple sexual partners in the last two months were found to be the most infected. Comparing the two methods, real-time PCR had a higher sensitivity (100%) and specificity (97.40%). This study shows that the prevalence rate of T. vaginalis infection remains low. However, using the PCR approach allows for better detecting infection than conventional staining method.