The study introduces the Cepheid GeneXpert MTB/RIF assay, a rapid, hands-free, near-patient technology for detecting *Mycobacterium tuberculosis* and identifying rifampin resistance. The assay uses a single-use cartridge with integrated sample processing and real-time PCR, capable of detecting as few as 4.5 genomes of *M. tuberculosis* per reaction. Analytical tests demonstrated a limit of detection (LOD) of 4.5 genomes per reaction, and clinical validation studies showed high sensitivity and specificity in detecting *M. tuberculosis* and rifampin resistance in sputum samples from patients in Vietnam and Uganda. The assay was also effective in detecting rifampin resistance in retreatment tuberculosis cases, with a specificity of 100% after correcting for conventional susceptibility test errors. The GeneXpert MTB/RIF assay offers a highly sensitive, simple, and rapid method for diagnosing tuberculosis and identifying drug resistance, which can significantly improve the management of tuberculosis cases, particularly in resource-limited settings.The study introduces the Cepheid GeneXpert MTB/RIF assay, a rapid, hands-free, near-patient technology for detecting *Mycobacterium tuberculosis* and identifying rifampin resistance. The assay uses a single-use cartridge with integrated sample processing and real-time PCR, capable of detecting as few as 4.5 genomes of *M. tuberculosis* per reaction. Analytical tests demonstrated a limit of detection (LOD) of 4.5 genomes per reaction, and clinical validation studies showed high sensitivity and specificity in detecting *M. tuberculosis* and rifampin resistance in sputum samples from patients in Vietnam and Uganda. The assay was also effective in detecting rifampin resistance in retreatment tuberculosis cases, with a specificity of 100% after correcting for conventional susceptibility test errors. The GeneXpert MTB/RIF assay offers a highly sensitive, simple, and rapid method for diagnosing tuberculosis and identifying drug resistance, which can significantly improve the management of tuberculosis cases, particularly in resource-limited settings.