Jordan has a low prevalence of the Human Immunodeficiency Virus (HIV)/Acquired Immunodeficiency Syndrome (AIDS). However, HIV/AIDS is an important global public health issue and medical students in Jordan need to be well-informed about it. To assess Mutah Medical School students’ knowledge about HIV/AIDS and to explore their attitudes toward people living with HIV (PLWH). A cross-sectional study conducted in 2013 among clinical years students using a self-administered questionnaire. A total of 209 students filled the questionnaire. Slightly more than half of the students reported having received adequate or more than adequate teaching on sterilization procedures (53%) and barriers (55%), while only 43% reported similar teaching received of cross-infection precautions. The students showed a good overall knowledge about HIV/AIDS and the transmission routes of HIV. However, some misconceptions existed such as thinking that HIV could be transmitted by mosquitos (30%) and flies (10%). Negative attitudes were displayed toward patients with HIV/AIDS with 89% agreeing that AIDS makes their job a high-risk occupation and 60% agreeing that given a choice, they would prefer not to work with AIDS patients.  Although there was a good overall knowledge of the students about HIV/AIDS, negative attitudes were prevalent towards PLWH.