Background: To reduce the burden of human immunodeficiency virus (HIV), the popularization of voluntary counseling and testing (VCT) for early determination of human immunodeficiency virus status will be of immense benefit. Aim: To evaluate the uptake of voluntary counseling and testing and sero-prevalence of human immunodeficiency virus among status naïve outpatients and self-presenting VCT clients in Benin City, Nigeria. Materials and Methods: This study was carried out in the period of May 2010 to April 2011 at the University of Benin teaching hospital, Benin City. Subjects who consented were included and screened for HIV. Pre and post-test counseling was done following the world health organization guidelines. Results: Out of 10,533 subjects (7783 outpatients and 2750 self-presenting VCT clients) counseled for VCT, a total of 4651(44.2%) subjects; (3971(51.0%) outpatients and 680 (24.7%) self-presenting VCT clients), consented and accepted HIV VCT. Overall HIV prevalence was 6.4%. 270 (6.8%) outpatients and 29 (4.3%) self-presenting VCT clients were HIV positive. HIV was significantly associated with female gender among the outpatients (P<0.001). Conclusion: VCT uptake was low; the sero-prevalence of HIV was high. The need to employ an expanded and more purpose oriented public enlightenment campaign on the usefulness of HIV VCT should be a priority for HIV control agencies in our area.