Aim: The present study aimed to test the validity, reliability and sensitivity of a two-question depression screening instrument with the Center for Epidemiologic Studies Depression Scale in human immunodeficiency virus-infected subjects. Methods: The present study is a self-administered questionnaire of the two-question depression screening instrument and the Center for Epidemiologic Studies Depression Scale-D10 completed contemporaneously by participants to assess the validity, reliability and sensitivity of the instrument. Results: Eighty participants (76 men and 4 women) with a mean age of 43.4years were enrolled to the study. In this cohort the prevalence of depression was found to be 36%. A significant correlation (rho= 0.68, P < 0.0001) was found between the depression screening instrument and Center for Epidemiologic Studies Depression Scale indicating adequate validity. The internal reliability of the depression screening instrument was found to be 0.77. The two-question instrument yielded a sensitivity of 89.7% and a specificity of 58.8%. The false positive rate was 41.2%. However, the false negative rate in this population was found to be only 10.3%. The negative predictive value was 90.9%. Conclusions: The results of the present study indicated that the two-question depression screening instrument was valid, reliable and sensitive for detecting individuals at risk of depression in a cohort of people living with human immunodeficiency virus attending a nutrition clinic in an enhanced care setting.