Controlled trials have demonstrated that liquid media culture (LMC) is superior to solid media culture for diagnosis of Mycobacterium tuberculosis (MTB), but there is limited evidence about its performance in resource-limited settings. We evaluated the performance of LMC in a demonstration project in Bangkok, Thailand. Sputum specimens from persons with suspected or clinically diagnosed tuberculosis were inoculated in parallel on solid (Lowenstein-Jensen [LJ]) and liquid (mycobacterial growth indicator tube [MGIT 960]) media. Biochemical tests identified isolates as MTB or nontuberculosis mycobacteria (NTM). Of 2566 specimens received from October 2004 to September 2006, 1355 (53%) were culture positive by MGIT compared with 1013 (39%) by LJ. Median time to growth for MGIT was significantly less than LJ: 11 versus 27 days. Of 1417 isolates detected by at least 1 media, 1255 (86%) were identified as MTB and 162 (11%) NTM. MGIT improved speed and sensitivity of MTB isolation and drug susceptibility testing, regardless of HIV status.