More than 1,000,000 persons in the United States are living with human immunodeficiency virus (HIV)/acquired immune deficiency syndrome, with 24% unaware of their HIV status. In this study, the authors explored patients' attitudes toward HIV testing in academic medical clinics and investigated the possible impact of the 2006 Centers for Disease Control and Prevention (CDC) HIV screening guidelines. Method: Cross-sectional survey study of adult patients in 9 academic internal medicine clinics (response rate 73%). The survey consisted of 76 questions, which assessed demographics, HIV risk factors, knowledge, beliefs, attitudes and characteristics of patient-physician interactions. Patient self-reported HIV testing was the main outcome. Bivariate analyses were performed, and variables with a P-value of <0.1 were included in a logistic regression model to determine characteristics most associated with HIV testing. Results: Four hundred forty-three patients completed the survey (response rate 73%) and 61% reported being screened for HIV. Physician recommendation (P < 0.0001), patient's own request (P < 0.0001), African American race (P < 0.0001) better knowledge about HIV (P = 0.0002), agreement with CDC recommendations (P < 0.0001), being comfortable with their doctor (P < 0.0001) and using street drugs (P < 0.0001) were all strongly associated with testing. In logistic regression, the only factors that remained statistically significant predictors of patients self-reported HIV testing were a patient's request for testing (OR: 103.3) and patient's knowledge about HIV (OR: 1.3). Conclusion: In this study, patient request was the strongest predictor for HIV screening and majority of patients accepted the idea of HIV testing in congruence with the CDC recommendations. Therefore, simple waiting room prompts and public education campaigns may represent the most efficient interventions to increase HIV testing rate. © Copyright 2010 Southern Society for Clinical Investigation.