Abstract: Objective：To compare the accuracy of different methods for identification of rapidly growing mycobacteria (RGM), and to detect the sensitivity of RGM isolates to anti-tuberculosis drugs. Methods：18 strains of RGM were identified by DNA microarray and gene sequencing method. The sensitivity for the RGM isolates of 9 anti-tuberculosis drugs, i.e., rifampicin, isoniazid, streptomycin, ethambutol, protionamide, aminosalicylic acid, capreomycin, ofloxacin and kanamycin, were tested by proportion method. Results：The coincidence rate for the identification of DNA microarray and gene sequencing to Mycobacterium fortuitum and Mycobacterium abscessus was 100%, but obvious deviation of identification results for the other species was found. RGM isolates were commonly resistant to the tested 9 anti-tuberculosis drugs. The resistant degree was as below: Mycobacterium massiliense (all resistant to 9 drugs), Mycobacterium abscessus (resistant to 8 9 drugs), Mycobacterium neoaurum (resistant to 8 antibiotics), Mycobacterium fortuitum (resistant to 7 drugs), Mycobacterium septicum (resistant to 7 drugs), Mycobacterium conceptionense (resistant to 6 grugs) and Mycobacterium phocaicum (resistant to 4 drugs). Conclusion：Molecular diagnostic technique should be of value in clinical laboratory. The treatment of anti-tuberculosis drugs for RGM infection may be greatly risky. The drugs breakpoints of rapid growing Mycobacterium should be adopted to guide the clinical application of anti-tuberculosis drugs.