Objectives: This cross-sectional study aimed to assess the knowledge and practice of universal precautions among Faculty of Medicine, Kuwait University, medical students in their clinical years. Subjects and Methods: A self-administered questionnaire including 9 knowledge and 9 practice questions of universal precautions was used. A score was calculated for both knowledge and practice. These scores were dichotomized into less than median score (poor) and more than median score (good). 244 students were invited to participate. Results: Of the 244 students, 220 (90.2%) accepted to participate. Of the 9 knowledge questions about universal precautions, 7 were answered correctly by more than 60% (range 64.8–90.4) of the students. All questions regarding the practice of universal precautions were answered correctly by over 60% (range 61.8–96.4) of the students. The prevalence of poor knowledge was 38.2% (84/220) and poor practice was 27.7% (61/220). Grade point average was significantly (p = 0.008) associated with the knowledge status of the respondents; however, it was not significantly associated (p = 0.397) with practice of universal precautions. Furthermore, neither the knowledge status nor any of the sociodemographic variables were significantly associated with the practice of universal precautions. Conclusions: A substantial number of students of the Faculty of Medicine, Kuwait University, in their clinical years appeared to have poor level of both knowledge and practice of universal precautions. Efforts are needed to optimize the level of knowledge and practice among students to minimize the risk of preventable infections.