Hepatitis B viral infection is a silent deadly disease. According to the Society for Gastroenterology and Hepatology in Nigeria (SOGHIN), over 20 million persons are infected with hepatitis and death from it is on a large scale. This study was carried out in order to estimate the prevalence of Hepatitis B Virus (HBV) among students and non-teaching staff of Nile University of Nigeria, Abuja. The study was cross-sectional; involving 200 participants (150 students and 50 non-teaching staff) recruited over a period of three months, from the first week of February 2017 to the end of May 2017 and screened for Hepatitis B surface antigen (HBsAg) using One Step Rapid Diagnostic Test. Structured questionnaires were also issued out to the study population. The study revealed that 13 out of the 200... people tested were positive giving an overall prevalence rate of 6.5% (95% CI: 3.0, 10.0%). 3 out of the 150 students were positive and 10 out of the 50 non-teaching staff tested were positive. The prevalence of HBsAg was higher among the non-teaching staff (20.0%) than among students (2.0%). Disaggregated by gender, the prevalence was higher among males (8.9%) than among females (4.0%). The reasons for the difference in prevalence and health implications of these findings are discussed.