ObjectiveTo study the species and drug resistance of Mycobacterium isolated from patients with sputum smear positive for acidfast bacillus in Wujiang city, and provide reference for the prevention and control of tuberculosis. MethodsSputum specimens with positive smear were cultured, isolated bacteria were identified and performed drug susceptibility testing, drug resistance among different species of strains and between patients with initial and repeated treatment were compared.ResultsA total of 103 Mycobacterium isolates were included in the study,13 of which were nontuberculous Mycobacterium, drug resistance rate was 100.00%, multidrug resistance (MDR) rate was 84.62%; 90 isolates were Mycobacterium tuberculosis, 81(90.00%) of which were Mycobacterium hominis. Drug resistance rate of Mycobacterium tuberculosis was 35.56%, MDR rate was 14.44%. Of 70 initially treated tuberculosis patients with positive sputum smear,14(20.00%) were resistant to drugs, MDR rate was 4.28%(3/70); Of 20 repeatedly treated tuberculosis patients with positive sputum smear,18(90.00%)were resistant to drugs, MDR rate was 50.00%(10/20).ConclusionMycobacterium tuberculosis is the major isolated strain from patients with positive sputum smear. Drug resistance and MDR rates of nontuberculous Mycobacterium are very high. Drug resistance and MDR rates of Mycobacterium tuberculosis in repeatedly treated patients are higher than initially treated patients.