Trichomoniasis caused by Trichomonas vaginalis is the most prevalent non- viral sexually transmitted disease. InPouch™ TV culture is highly sensitive in diagnosing the disease. Unfortunately, researches made in Libya to study the prevalence of trichomoniasis are very few; also diagnosis is based only on clinical manifestations which could be confused with those of other sexually transmitted diseases. Hence, the present work was aimed to determine the prevalence of Trichomonas vaginalis infection in women suffering from vaginal discharge in Zawia district, Libya, also to compare between InPouch™ TV culture, wet mount examination and Giemsa staining for the diagnosis of trichomoniasis. It was found that infection rate was relatively high in the examined patients. InPouch™ TV culture was the most sensitive method and detect 36.7% positive patients in the examined area. Also, its specificity was 100%. It is concluded that infection rate of Trichomonas vaginalis in Libya is relatively high, so there must be a policy decision to increase awareness of this disease. It is recommended to use this culture as a routine method for diagnosis in the laboratory.