Lymphatic filariasis is a significant public health and economic problem in many tropical and sub-tropical regions. Unplanned urbanization leading to a lack of proper sanitary conditions has resulted in an increase in the urban-based transmission of a number of vector-borne diseases, including lymphatic filariasis. It has been well established that lymphatic filariasis is endemic in rural areas of Ghana. The goal of this study was to determine if there is a potential of establishing urban transmission cycles in Ghana's major cities. We clinically and immunologically assessed 625 individuals from the three major urban areas (Bawku, Bolgatanga and Secondi/Takoradi), finding that the prevalence of infection with Wuchereria bancrofti ranged from 0 to 12.5%. The results of a polymerase chain reaction based analysis of mosquitoes collected from these areas suggested that there is a low but detectable prevalence of mosquitoes infected with W. bancrofti. We conclude that there may be a potential for an established urban transmission of lymphatic filariasis in Ghana.