Based on the estimation of World Health Organization (WHO), there were 558000 new cases with resistance to Rifampicin, of which 82% had Multidrug-resistant Tuberculosis (MDR-TB). We aimed to identify the outbreakof MDR-TB in River Nile state, Sudan, and the risk factors contributing to its occurrence. This descriptive cross-sectional hospital-based study involved 200 specimens from patients suspected of having MDR-TB tested using an automated GeneXpert assay. Theresults of the GeneXpert assay showed that the presence of Mycobacterium tuberculosis in 81 (40.5%), and out of 81 positive test results there were 13 (16%) had MDR-TB. Additionally, 7 cases of MDR-TB were previously treated, which represented about (53%) of MDR patients. The remaining 6 MDR-TB patients were new cases and represented (47%) of MDR-TB patients. Moreover, 4 MDR-TB patients had a backgroundof contact with MDR-TB patients. Prevalence of MDR-TB in River Nile State, Sudan was 16%, which is greater than WHO estimation for Sudan (10.1%). The results revealed that the leadingrisk factor in developing MDR-TB was a backgroundof contact with MDR-TB, so adherence to treatment and social awareness about the spread of MDR-TB isa crucial preventive measure.