Aim: Determine the prevalence of serological markers and identify risk factors associated with Hepatitis B Virus (HBV) infection in patients screened at the Bouake teaching hospital. Study Design: Retrospective cross sectional study was conducted Study Site and Period: Bacteriology-Virology Laboratory/Bouake teaching hospital, Côte d´Ivoire, from April 2016 to January 2018. Methodology: In all 1076 study participants, venous blood sample was collected and screened for HBV surface antigen (HBsAg) and antibody against HBV core antigen (anti-HBc), by electrochemical-luminescence following the manufacturer protocols.  Additionally, questionnaires were used to collect information regarding sociodemographic variables and possible risk factors for hepatitis B infection.  Data were processed and analyzed using EPI INFO 7 software. Results: A total of 1076 participants were included in this study with a median age of 30.0 years (range: 3 months; 82 years). Of which, 514 (48%) were female and 562 (52%) were male with female / male ratio 1.09. HBsAg was detected in 24,3% of participants and 82 1 (76,3%) were exposed to the risk of HBV infection (anti-HBc positive). High rate of HBV infection was detected in male (27.93%) (p=0.003). The age group of 15–45 years were more infected (27.18%) (p<0.0001). The detection rate of HBe antigen (HBeAg), anti-HBe and anti-HBc (total antibodies) were respectively 12%; 86% and 7%. Of 938 participants who were not vaccinated against HBV, 240 (25.58%) were HBsAg-positive. HBV vaccine uptake was protective against HBV infection (AOR =0.580; 95% CI 0.359-0.938; p=0.024). Conclusion: The rate of carriage of HBs antigen was higer than national rate, which confirms that Bouake is a highly endemic area for HBV infection. Vaccination against Hepatitis B virus is the only way to prevent and to fight effectively against this infection. It is therefore important to encourage the screening and vaccination in the general population.