University students in Ghana, as in elsewhere in Africa are reported to be engaging in lifestyles that put them at risk of HIV infection. This study was conducted to examine possible gender differences in the knowledge, attitudes, and behaviors related to HIV among university students. A self-administered questionnaire was used to collect data from 181 randomly selected Ghanaian university students. The results showed that over half (52%) of the participants were sexually active and had engaged in various sexual practices including unprotected sexual intercourse. About 63% had used condom in their previous sexual activity prior to the study and 76% indicated that they were intending to use condoms in the future. Approximately three-quarters (71%) of the participants did not perceive themselves at risk for HIV infection. Gender differences were observed in HIV knowledge, sexual risk behaviour and attitudes towards persons with HIV/AIDS. This research provides additional insights that may be used to develop effective HIV prevention strategies in Ghana.