Tuberculosis (TB) has reemerged as a major public health problem worldwide. Although traditional acid-fast smears and cultures remain important tools for laboratory diagnosis of TB, these methods are not adequate to meet the challenges of the current crisis. In response to urgent needs for more rapid and sensitive diagnostic methods in mycobacteriology, several innovative procedures have been developed. Radiometric, molecular, biochemical, and chromatographic techniques have been introduced successfully in clinical laboratories, and others continue to be investigated. I review the newer technologies that provide laboratories with the tools necessary to achieve rapid detection, identification, and susceptibility testing of Mycobacterium tuberculosis.