The paper aims to investigate the susceptibility test results of 201 tuberculosis patients in Shanghai Pulmonary Hospital from December 2008 to May 2009. Roche system was used to culture Mycobacterium tuberculosis (M. tuberculosis), and absolute concentration method was used to test drug susceptibility.  In 201 patients, the resistance rates of M. tuberculosis were as follows: isoniazid (INH, 52.24%), streptomycin (SM, 45.77%), rifampicin (RFP, 30.35%), ethambutol (EMB, 28.86%), amikacin (AK, 28.18%), rifapentine (RIB, 26.40%), pasiniazide (Pa, 25.56%), paraminosalicylic acid (PAS, 23.60%), protionamide (Pt, 23.03%), capreomycin (CPM, 20.56%). Among all the patiemts, drug resistance rates in initial treated patients was 3.08%~25.87%, in previously treated patients was 27.50%~64.71%, in multi-drug resistant patients was 64.44%~93.10%. Overall, compared with low concentration of drugs, high concentration of SM, INH, EMB, RIB and Pt decreased the resistance rate of M. tuberculosis significantly (P <0.05). Except for INH, SM and RFP, the most drug resistance rates and multi-drug resistance rate in female were significantly higher than those in male (P <0.05). The results suggest that the application of anti-tuberculosis drugs should be adequate clinically, and more attention should be paid to the female patients.