Background: Health-care workers (HCWs) are often exposed to potentially infectious body fluids-carrying Hepatitis B virus (HBV) from their workplaces. Typically, most asymptomatic HBV carriers-status may remain unnoticed for decades, despite a few percentages of them with vaccination history. In this study, we aimed at investigating asymptomatic carriage of HBV and its associated risk factors among co-health workers ultimately impacting on patients. Methods: Venous blood samples and demographic data were obtained from a cross sectional survey of HCWs categories working in OOUTH, a referral teaching hospital and other selected health facilities in Ogun state. Immuno-chromatographic discs and Enzyme Linked Immuno-Sorbent assay (ELISA) techniques were adopted for the determination and confirmation of the presence of HBsAg, anti-HBs and total anti-HBc in sera. A structured questionnaire was used for obtaining demographic data which were analyzed by regression analytical technique. Results: About 11.44% (43) of the 376 healthcare workers (HCWs), were infected with HBV. The serological marker of viral particles identified in this positive study population included Hepatitis B envelope antigen (53.49%), Hepatitis Core antigen (13.15%), Hepatitis B surface antibodies (3.99%); and Hepatitis B core antibodies (0.80%). A significant association existed between risk factors such as recapping of used needles and carriage of HBV among the HCWs (p-value =0.017). Conclusion: The carriage rate (11.44%) of HBV among HCWs in Ogun state referral health facilities as against the 8% set standard by W.H.O. in 2009, depict the associated risk factors of asymptomatic carriage, pathogenesis and dissemination in and outside of the health facilities.