Human immunodeficiency virus (HIV) and Hepatitis B virus (HBV) co-infection have been associated with reduced survival, increased risk of liver disease and hepatotoxicity despite undergoing antiretroviral therapy (ART). This study aims to determine the prevalence of serological markers of HBV based on serological markers among infected individuals with HIV. Ninety-three sera samples of HIV positive individuals were collected by Ladoke Akintola University of Technology (LAUTECH) Teaching Hospital-HIV/AIDS Supporting and Counselling Unit, Ogbomoso. The collected samples were analyzed for enumeration of CD4+ cells and presence of HBV. The selected three HBV serological markers were HBsAg, HBeAg and HBcAb while using a third generation ELISA kit for screening. The mean CD4+ count and age were 479.12 ± 23.48 cells/µl and 48.56 ± 1.27 years, respectively. Only six samples out of ninety-three HIV-positive samples were positive to HBsAg while fifty-four samples (58.1%) were positive for HBcAb however, HBeAg marker was negative among all samples. Interestingly, the prevalence of the HBV infection was more in females (66.6%) compared to the male counterparts (33.3%) within the overall screening. In conclusion, the current study describes the prevalence of HBV infection amongst HIV-positive individuals in Ogbomoso. The high prevalence of HBcAb confirmed that Ogbomoso district is highly endemic with HBV infection. Likewise, HBV vaccination in HIV-infected patients is encouraged with careful monitoring for liver function, particularly those under ART.