Yohannes Sinku,1,2 Takele Gezahegn,1 Yalewayiker Gashaw,1 Meseret Workineh,1 Tekalign Deressa1 1School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, 2Diagnostic Laboratory Case Team, University of Gondar Teaching Hospital, Gondar, Ethiopia Background: The epidemiology of human immunodeficiency virus (HIV) infection in Ethiopia varies with regions, study population, and time. Thus, timely information on HIV epidemiology is critical for the combat of the epidemic. In this study, we aim to update HIV prevalence and risk factors among voluntary counseling and testing (VCT) clients at the University of Gondar Teaching Hospital, Northwest Ethiopia.Methods: A total of 2,120 VCT clients’ records from September 2007 to August 2010 were analyzed retrospectively. Bivariate logistic regression analysis was used to identify significant predictors. Odds ratio (OR) and 95% confidence intervals (CIs) were calculated. Statistical significance was set at P-value <0.05.Results: Of 2,120 VCT clients, 363 (17.1%) were seropositive for HIV. A higher rate of HIV positivity was observed among female clients (20.4%) than that in male clients (14.0%) (OR 1.58, 95% CI 1.26–1.98, P=0.00). Widowed (95% CI 10.42–34.92, P=0.00), married (95% CI 3.42–5.94, P=0.00), divorced (95% CI 2.79–5.32, P=0.00), and illiterate (95% CI 2.33–5.47, P=0.00) clients were associated with HIV infection with the odds ratios of 19.07, 4.51, 3.85, and 3.57, respectively. Clients within the age category of 35–49 years (OR 5.03, 95% CI 3.56–7.12, P=0.00) and above the age of 50 years (OR 4.99, 95% CI 2.67–9.34, P=0.00) were more likely to be infected with HIV.Conclusion: HIV is still the major concern of public health in the Gondar area as evidenced by our data. Being female, widowed, married, illiterate, and older age were the identified risk factors for HIV infection. Thus, consideration of these factors in future intervention and clinical practice is recommended. Keywords: HIV, AIDS, voluntary counseling and testing, prevalence, Northwest Ethiopia