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According to the World Health Organization (WHO) estimates, there were 558,000 new cases of rifampicin-resistant tuberculosis, with 82% of these classified as multidrug-resistant tuberculosis (MDR-TB). This study aimed to investigate the outbreak of MDR-TB in River Nile State, Sudan, and identify the risk factors associated with its occurrence. A descriptive cross-sectional hospital-based study was conducted involving two hundred specimens from patients suspected of having MDR-TB, tested using the automated GeneXpert assay. The GeneXpert results indicated that Mycobacterium tuberculosis was detected in 81 cases (40.5%), and among these positive results, 13 (16%) were confirmed as MDR-TB. Furthermore, 7 of the MDR-TB cases had a history of previous treatment, accounting for approximately 53 percent of MDR patients. In contrast, the other 6 MDR-TB cases were new, representing 47% of the MDR-TB patients. In addition, 4 MDR-TB patients had a history of contact with other MDR-TB cases. The prevalence of MDR-TB in River Nile State, Sudan, was found to be 16%, which is higher than the WHO estimate for Sudan at 10.1%. The findings highlighted that prior contact with MDR-TB patients is the primary risk factor for developing MDR-TB, emphasizing that treatment adherence and increased social awareness about MDR-TB transmission are essential preventive measures.