Anti-tuberculosis drug-induced hepatotoxicity is a common serious adverse drug reaction. This study intended to determine the prevalence and associated factors of drug-induced hepatotoxicity among tuberculosis and human immunodeficiency virus co-infected patients in Dessie referral hospital northeast Ethiopia.In this cross-sectional study 84 patients were enrolled retrospectively. Data from September 1/2015 to August 30/2018 were extracted from March 1/2019 to April 1/2019. Association between dependent and independent variables was determined using the odds ratio and a P value of < 0.05 was considered as statistical significance. Out of 84 patients, 17 patients developed drug-induced hepatotoxicity which makes the prevalence of drug-induced hepatotoxicity 20.2%. The result revealed that the presence of disseminated or extrapulmonary tuberculosis [(AOR: 7.728, 95% CI (1.516-39.404)] and/or body mass index less than 18.5 kg/m [(AOR = 5.593, 95% CI (1.180-26.519)] were a risk factor for drug-induced hepatotoxicity. Tuberculosis and human immunodeficiency virus co-infected patients with extra- pulmonary tuberculosis and/or body mass index less than 18.5 kg/m2 should be closely followed and supervised for the development of hepatotoxicity.