Background <p>This study aimed to determine the prevalence of Tuberculosis (TB) and rifampicin-resistant Tuberculosis (RR-TB) among Sudanese patients tested using the GeneXpert assay.</p> Methods <p>Data were extracted from medical records of presumptive tuberculosis patients attending the hospital between February and October 2025. Sputum samples were analyzed using the GeneXpert MTB/RIF assay to detect <i>Mycobacterium tuberculosis</i> and rifampicin resistance.</p> Results <p>A total of 153 sputum samples were analyzed. male participants comprised 71.0%, and female participants 28.1%. The majority (41.8%) were aged 40–64 years, followed by 25–39 years (33.3%), 15–24 years (12.4%), and children aged 1–14 years (2%). <i>M. tuberculosis</i> was detected in 29 cases (19.0%), with 81% tested negative. Among positive cases, medium bacillary levels were most common (48.3%). Rifampicin resistance was detected in one case (0.7%), with one additional inconclusive result; the remaining positive cases were rifampicin sensitive (18.3%). A small proportion of participants reported previous TB Infection (4.6%) or HIV Infection (1.3%). Chi-square analysis revealed no statistically significant relationship between age (<i>p</i> = 0.454) or gender (<i>p</i> = 0.228) and TB infection. In contrast, previous TB Infection(<i>p</i> &lt; 0.001), and HIV Status (<i>p</i> = 0.018) were strongly associated with TB detection. No significant association was observed between age, gender, previous TB infection, and rifampicin resistance (<i>p</i> &gt; 0.05).</p> Conclusions <p>The findings suggest a high prevalence of tuberculosis and notable recurrence among presumptive cases, suggesting deficiencies in the treatment cascade. The low rate of rifampicin resistance is a cautiously optimistic, preliminary finding, highlighting the need for strengthened surveillance and continuity of care in Sudan’s conflict-affected setting.</p>