Trichomoniasis is the most common non-viral sexually transmitted infection, and it is caused by the protozoan flagellate Trichomonas vaginalis. Although highly prevalent in sexually active women, it has long been overlooked in other groups of potentially infected people. Recently, studies have shown that trichomoniasis increases the risk of infection with human immunodeficiency virus and can cause adverse outcomes of pregnancy, which has increased interest in T. vaginalis and increased the need for highly sensitive diagnostic tests. This article summarizes the diagnostic methods most commonly used in the diagnosis of trichomoniasis, including the most sensitive and specific nucleic acid amplification tests. It also presents the results of our study comparing the performance of wet mount microscopy and culture to real-time PCR for detecting the parasite.