Abstract: Objective To analyze the result of clinical isolates (strains of acid-fast bacilli) and sputum samples using Gene Xpert Mtb/RIF (Xpert) and to evaluate the value of Xpert.Methods One hundred and forty-eight clinical isolates and 40 sputum samples were detected using Xpert. Forty sputum samples were cultured with acid Lwenstein Jensen (L-J) me-dium. All clinical isolates were identified with medium (PNB) and performed drug susceptibility testing(DST).Results (1) 144 Mycobacterium tuberculosis(Mtb) and 4 nontuberculosis mycobacteria(NTM) were identified by Xpert in 148 clinical isolates, and 37 Mtb and 3 NTM were identified by Xpert in 40 sputum samples. The result was consistent of identification for 4 NTM strains by PNB and Xpert.(2) Culture was positive in 40 smear-positive sputum samples. 181 Mtb were identified in total 188 clinical isolates. Of 181 Mtb, rifampin resistance was 8 strains including 1 mono-resistant to rifampin,6 multidrug-resistant tuberculosis and 1 extensively drug resistant tuberculosis. The DST result was consistent of Xpert’s in strains resistant to rifampin.Conclusion Xpert can assist judgment Mtb and NTM. It has great value to find patient with tuberculosis and drug resistant tuberculosis.