Introduction. Tuberculosis is still a major worldwide health problem. It is responsible for the death of 3 million people each year. Epidemiological studies with techniques which allow differentiation of strains within the Mycobacterium tuberculosis (MT) are important to limit the dissemination of the disease. Therefore, there is a great need for an improved methods to subtype MT strains by a simple and rapid molecular finger-printing method by using double repetitive element-polymerase chain reaction (DRE-PCR) analysis, yields a unique, strain-specific pattern of bands.Materials and Methods. MT was obtained from the national research institute of Tuberculosis and lung diseases. To perform DRE-PCR, the DNA molecules were extracted, then implicated with PCR. implication products were analyzed by gel electrophoresis on 2% agarose and generated patterns compared with together. Those patterns that identical for two or more strains among their 70 MT, were considered as cluster patterns.Results.Fourty-two different patterns were observed in the 70 isolated cases and analyzed. The rate of diversity was 60%, which is the same (approved) as the other studies. Thirty-one (44.3%) of the specimens had unique pattern, and 39 (55.7%) of specimens had strains belonging to one of the 11 clusters. The patterns had 1-6 band them approved the other studies. There was no correlation found among the special patterns, resistance to antituberculosis agents, residence, migration, and Iranian patients. The results are the same as what is reported in the literature.Conclusion. The data show the DRE-PCR method is the simplest and the fastest way to achieve molecular typing of MT in Iran.