Few studies have examined the vitamin D status in HIV-infected patients. A cross-sectional retrospective chart review of 2992 HIV-infected patients was conducted from 9/2008 to 5/2009. A total of 274 adult patients had 25-hydroxyvitamin D [25(OH)D] obtained by radioimmunoassay. None was receiving vitamin D (vitD) supplements. Vitamin D status was defined as the following: vitD deficiency (vitDd) as 25(OH)D <25 nmol/liter, vitD insufficiency (vitDi) as 25(OH)D 25-74 nmol/liter, and vitD optimal (vitDo) as 25(OH)D ≥75 nmol/liter. We analyzed demographic/laboratory data. vitDd, vitDi, and vitDo were 21.2% (58 patients, 58/274), 68.6% (188 patients, 188/274), and 10.2% (28 patients, 28/274), respectively. There were significant racial differences. Blacks were 60.3% (35 patients, 35/58), 40.4% (76 patients, 76/188), and 28.6 % (8 patients, 8/28) in vitDd, vitDi, and vitDo, respectively, p=0.002. CD4 T cell count was not different in these three groups. However, HIV viral load was significantly different. Median log (10) HIV viral load was 2.31 with IQR 1.70-409, 1.70 with IQR 1.70-2.96, and 1.70 with IQR 1.70-2.78 in vitDd, vitDi, and vitDo, respectively, p=0.039. Multivariate logistic regression analysis showed that black race [odd ratio (OR) 4.108, 95% confidence interval (CI) 1.462-11.543, p=0.007] and HIV viral load>50 copies/ml (OR 2.396, 95% CI 1.120-5.127, p=0.024) were significantly associated with vitamin D deficiency. Vitamin D deficiency was highly prevalent in HIV-infected patients. Detectable HIV viremia and dark skin (black ethnicity) were significantly associated with vitamin D deficiency. Evaluation of vitamin D status in HIV-infected patients should be considered and further studies are needed to define the effects of vitamin D.