Introduction Hepatitis B virus (HBV) infection is hyper-endemic in Nigeria. Prisons are high-risk environments for the spread of infectious diseases. Worldwide, seroprevalence of HBV infection is substantially higher among individuals in correctional facilities when compared to general population. We determined the seroprevalence and risk factors associated with HBV infection among Kuje prison inmates, Nigeria. Material and methods We conducted a prison facility based cross-sectional study. Interviewer administered questionnaires were used to obtain information on participants socio-demographic characteristics, HBV risk factors, previous HBV test and vaccination history. Blood samples collected from participants were analysed for HBsAg, HBsAb, HBcAb, HBeAg and HBeAb markers using rapid lateral chromatographic immunoassay kit. Univariate, bivariate, and multivariate analysis were performed. Results A total of 271 inmates (63 convicts and 208 awaiting trial inmates) were recruited into the study as participants. The mean age of the participants was 32.7 SD±9 years. HBV seroprevalence (HBsAg) of 13.7% (95% CI: 9.8-18.3) was found. 55.4% (95% CI: 49.2-61.4) of inmates were susceptible to HBV infection, 20.7% (95%CI; 16.0-26.0) had past HBV infection while 10.3% (95% CI: 7.0-14.6) had acquired natural or artificial HBV immunity. Factors found to be associated with current HBV infection (HBsAg) include age-group [less than or equal to]25years (aOR = 8.0,95% CI: 2.9-22.3), being ever married (aOR = 4.2, 95% CI: 1.7-10.4) and history of alcohol consumption (aOR = 3.4, 95% CI: 1.3-8.4). Conclusion This study reveals a high seroprevalence of HBV infection among Kuje Prison inmates, hence the need to introduce prison-focused health intervention initiatives such as HBV screening, vaccination and care to reduce the transmission of HBV infection among inmates and ultimately the general population.