Abstract Background Hepatitis B virus (HBV) and tuberculosis (TB) represent major public health problems. There is currently little data on HBV infection among TB patients with and without human immunodeficiency virus (HIV). Objectives To assess HBV prevalence among TB patients with and without HIV. Study design From April 2008 to March 2010, a cross-sectional study was conduct among TB patients attended at a reference hospital in Goiânia City, Brazil. The participants were tested for serological markers of HBV infection and HIV antibodies. HBV DNA was detected in HBsAg-positive samples, and also in HBsAg-negative/anti-HBc-positive samples to look for HBV occult infection. Results Of 425 patients, 402 (94.6%) agreed to participate in the study. The overall prevalence of HBV (HBsAg and/or anti-HBc positive) and HIV infections were 25.6% (103/402) and 27.6% (111/402), respectively. A higher HBV infection rate was found among HIV-infected patients (36.9%; 41/111) compared to patients infected with TB only (20.0%; 57/285). A multivariate analysis of risk factors showed that age ≥50 years ( p = 0.03), non-injecting ( p < 0.01) and injecting ( p < 0.01) drugs use were associated with HBV infection. Among the HBsAg-positive samples ( n = 13), HBV DNA was detected in 10 (76.9%) samples. Of the 90 anti-HBc-positive samples, 13 were HBV DNA positive (with very low levels) resulting in an occult HBV infection rate of 14.4%. PCR-RFLP was successfully performed in 20 HBV DNA-positive samples: 15 were genotype A and 5 were genotype D. Conclusions HBV infection was common, particularly among this with HIV infection.