Objectives: To clarify the awareness and source of AIDS knowledge among college students of medical university of Guizhou province, China, so as to provide some basic evidences for implementing education of AIDS control and prevention. Methods: A stratified cluster sampling survey was conducted. Self-designed questionnaires were used to investigate university students, Epidata 3.0 and SPSS 18.0 were used to analyze data. Results: The awareness rate of AIDS through blood transmission and mother-to-child transmission was over 90%, the awareness rate of sexual transmission was just about 80%, and the awareness rate of non-transmission route "mosquito bites will not spread AIDS" was only 53.3%. The awareness rate of AIDS transmission and treatment among medical students was higher than that of non-medical students. The Internet, propaganda materials, newspapers and magazines, and school classes were the main ways for university students to acquire AIDS knowledge. Some students hope to acquire knowledge from classmates and friends, it was found that the proportion of medical students receiving AIDS knowledge from school classes was significantly higher than that of non-medical students (80.8% vs. 73.1%, p = .007). Conclusion: Though the awareness rate of AIDS knowledge among medical students is better than that of non-medical students, they had insufficient knowledge of AIDS, especially the sexual transmission. Moreover, school classes may be an effective way to propagandize AIDS-related knowledge, so schools can consider publicizing the AIDS education through classes, so as to improve students' abilities of AIDS control and prevention.