Objective To analyze influence mycobacterium tuberculosis sputum culture and heat-proof rifabutin antimicrobial susceptibility test results and related factors.Methods To the centers for disease control and prevention of Huainan tuberculosis clinic into the program to treat 227 sputum smear-positive patients in the sputum culture and heat-proof rifabutin of antimicrobial susceptibility test. Results Among 227 patients sputum culture positive 220 cases, accounting for 96. 9％, sputum culture-negative 2 cases, accounting for 0. 9％, pollution, 5 cases of 2. 2％. Resistance rifabutin antimicrobial susceptibility test,a new patient resistance ratio for 10. 4％ ,early cure loser for 37.5％, concomitant person for 28.9％. Conclusion The mycobacterium tuberculosis in the process of cultivation should be strictly according to the requirements of collecting, processing, specimen vaccination, preservation and observation. Early cure failure patients and treat patients resistant rifabutin rate was far higher than new patients, the patients with irregular or unreasonable chemotherapy were concerned.