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To validate the Hopkins Symptom Checklist-25 (HSCL-25) for use as a depression screen amongst human immuno-deficiency virus (HIV) positive pregnant women.Amongst 903 (mean age 24.8 years) HIV-positive pregnant women, a two-phased design included measures for health-related quality of life, perceived social support, and the HSCL-25 screen for depressive (HSCL-15 subscale) and anxiety symptoms. The Structured Clinical Interview for DSM-IV (SCID) was independently administered on a stratified random subsample.Internal consistency of the HSCL-25 (alpha 0.93) and HSCL-15 (alpha 0.9) was adequate, with expected findings demonstrated in discriminant validity analysis. A depression-anxiety construct explained nearly 40% of the variance. Eight individual HSCL-25 items demonstrated an area under the curve (AUC) greater than 0.6 for DSM-IV major depression and the HSCL-25 and HSCL-revised had an optimal depression cut-off score of 1.06 and 1.03 for the HSCL-15.The HSCL-25 demonstrated utility as a screen for depression; its inability to gauge severity of symptoms in this cultural context is discussed.