Hepatitis B virus (HBV) and tuberculosis (TB) represent major public health problems. There are limited data on HBV infection among TB patients without Human Immunodeficiency Virus (HIV). The goal of this study was to assess the prevalence of Hepatitis B virus infection among tuberculosis patients. This was a cross-sectional study conducted among TB patients attending clinic at TB centres in Ebonyi State (Mile 4 Hospital and Mater Misericordiae Hospital). Questionnaire was used in obtaining information on the demographic and social life pattern of the patients. Blood samples were collected from the patients, EDTA – anticoagulated blood samples were used for CD4 T-cells estimation, while the serum extracted from the clotted samples was used for HBsAg, HBeAg and ALT estimation. A total of 145 TB patients were studied with 12 (8.3%) and 1(0.7%) being seropositive to HBsAg and HBeAg respectively. The only patient that was seropositive to HBeAg had an elevated Alanine transaminase (ALT) of 147mg/dl and CD4 T-cells count of 584cells/µl. Of the 145 subjects, 83(57.2%) were on Directly Observed Treatment Scheme (p=0.449) while 62(42.8%) were not (p=0.597). 6(4.2%) of the patients that were on DOTS were seropositive to HBsAg. A univariate analysis of risk factors showed that none of the risks were associated with HBV. The ALT values for those on DOTS showed that elevated ALT values were higher than the normal values with 6(4.2%) and 2(1.4%) seropositive to HBsAg. Considering the ALT values of patients yet to initiate DOTS, those with normal values showed higher seropositivity of 4(2.8%). For patients that had not started DOTS, also those with CD4 T-cells count of ≥500cells/µl demonstrated the highest seropositivity of 3.5% to HBsAg with mean value of 703.4±313.3. This study demonstrated a low prevalence of Hepatitis B virus infection among tuberculosis patients. HBV co-infection on tuberculosis patients on DOTS poses synergistic increase in hepatotoxicity among patients.