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Mood and anxiety disorders, particularly depression, and substance abuse (SA) commonly co-occur with HIV infection. Appropriate policy and program planning require accurate prevalence estimates. Yet most estimates are based on screening instruments, which are likely to overstate true prevalence.Large academic medical center in Southeast.A total of 1,125 patients, representing 80% of HIV-positive patients seen over a 2.5-year period, completed the Substance Abuse-Mental Illness Symptoms Screener, a brief screening instrument for probable mood, anxiety, and SA disorders. Separately, 148 participants in a validation study completed the Substance Abuse-Mental Illness Symptoms Screener and a reference standard diagnostic tool, the Structured Clinical Interview for DSM-IV.Using the validation study sample, we developed logistic regression models to predict any Structured Clinical Interview for DSM-IV mood/anxiety disorder, any SA, and certain specific diagnoses. Explanatory variables included sociodemographic and clinical information and responses to Substance Abuse-Mental Illness Symptoms Screener questions. We applied coefficients from these models to the full clinic sample to obtain 12-month clinic-wide diagnosis prevalence estimates.We estimate that in the preceding year, 39% of clinic patients had a mood/anxiety diagnosis and 21% had an SA diagnosis, including 8% with both. Of patients with a mood/anxiety diagnosis, 76% had clinically relevant depression and 11% had posttraumatic stress disorder.The burden of psychiatric disorders in this mixed urban and rural clinic population in the southeastern United States is comparable to that reported from other HIV-positive populations and significantly exceeds general population estimates. Because psychiatric disorders have important implications for clinical management of HIV/AIDS, these results suggest the potential benefit of routine integration of mental health identification and treatment into HIV service sites.