Abstract: Objective To understand the characteristics of gene mutation in four first-line anti tuberculosis drugs. Methods Mycobacterium tuberculosis strains were isolated from sputum samples of 19 MDR-TB infected patients and 254 sensitive TB infected patients for drug sensitivity tests. PCR amplification of DNA extracted from Mycobacterium tuberculosis was performed. The amplified products were sequenced and compared with the standard strain H37Rv. Results 19 strains of MDR-TB Mycobacterium and 254 strains of all sensitive TB Mycobacterium were not detected in isoniazid resistant inhA gene mutation. Multi drug resistant group mutation rate was the highest in isoniazid resistant gene katG, and mutation site of katG gene was 315. Rifampin resistant mutant gene was rpoB.RpoB resistant gene loci were 526 and 531, of which the mutation rate of 531 was higher than that of 526. Mutation of ethambutol resistance was embB, and embB gene resistance locus was 206. Streptomycin resistant mutant gene were rpsL and rrs, and the resistance locus of rpsL gene was 43, and the resistance locus of RRS gene was 1401, and the mutation rate of rpsL gene was higher than that of rrs gene. Conclusions katG, rpoB and embB, rpsL and rrs genes account for isoniazid, rifampicin, ethambutol and streptomycin multi-drug resistant mutation of mycobacterium tuberculosis. This study provides a theoretical basis for rapid diagnosis and control of multi drug resistant TB in the Urumqi city.