Objective: Human immunodeficiency virus (HIV) remains a critical public health challenge and disproportionately affects young adults, racial minorities, and residents of the Southern United States. This study aimed to investigate HIV knowledge, risk perception, and testing behaviors among a sample of college students in South Carolina (SC). Participants: Undergraduate students (N = 256) from a large public university in SC. Methods: Participants completed a cross-sectional survey on HIV knowledge, testing behaviors, and attitudes. Results: More than one-third of participants displayed misunderstanding about HIV transmission. Less than 20% perceived themselves to be at-risk for HIV, and only 8% reported having ever been tested for HIV. In addition, a majority of participants (83%) reported they would feel ashamed if they acquired HIV. HIV knowledge was negatively correlated with HIV testing. Conclusions: Results indicate the need for HIV education among college students in SC to clear misconceptions, minimize knowledge gaps, reduce HIV-related stigma, and promote HIV testing.