Mycobacterium tuberculosis remains one of the most significant morbidity and mortality. The rapid diagnosis of tuberculosis and detection of Rifampin (RIF) resistance are essential for early disease management. The GeneXpert MTB/RIF assay is a novel integrated diagnostic device for the diagnosis of tuberculosis and rapid detection of RIF resistance in clinical specimens. We determined the performance of the MTB/RIF assay for rapid diagnosis of tuberculosis and detection of rifampin resistance in smear-positive and smear-negative pulmonary and extra pulmonary specimens obtained from possible tuberculosis patients.