Objective To evaluate the application of multiple locus variable number tandem repeat analysis (MLVA, 15-locus set)for genotyping of Mycobacterium tuberculosis (MTB) strains of Beijing genotype. Methods Total 72 Beijing genotype MTB strains obtained from Beijing Thoracic Hospital were genotyped by MLVA (15-locus set). The results were compared with that generated from gold standardIS6110-RFLP. Results After genotyped by MLVA ( 15-locus set), 72 strains were grouped into 59 types,of which 53 were unique types. The Hunter-Gaston index (HGI) of MLVA ( 15-locus set) was 0.990. The loci QUB-11b, Mtub 21 and QUB-26 were polymorphic in selected Beijing genotype strains. Genotyping by IS6110-RFLP generated 69 types, of which 66 were unique types. The HGI of IS6110-RFLP was up to 0.999, and the MLVA (15-locus set) clustered strains could be further subdivided. Conclusion MLVA(15-locus set) showed better discriminatory ability in Beijing genotype MTB strains, though secondary typing of clustered strains by IS6110-RFLP is needed. Key words: Myeobacterium tuberculosis ; Minisatellite repeats; Genotype; Polymorphism,restriction fragment length