Tuberculosis (TB) is an infectious disease the second highest cause of death worldwide after HIV. The World Health Organization in Annual Report on Global TB Control 2015 is still set Indonesia, among 22 other countries (highburden countries), in third place as the country with people with Tuberculosis (TB) is the highest after India and China. The purpose of this study was to detect rpoB gene as a marker of resistant strains of MDR TB in Papua by PCR. From 81 sempel in get mikrroskopis Negative results: 64 samples, Scanty: 2 samples, Positive 1: 3 Sample, Positive 2: 4 and Positive Sample 3: 8 Samples. Of the 19 positive samples of microscopic done rpoB gene identification method GenXpert in get 100% positive and are resistent to the antibiotic rifampicin. Of the 19 positive samples of microscopic test and test GeneXpert proceed to Culture as a gold standard method and obtained 100% positive. And antibiotic resistance test terhadap4 obtained the following results: antibiotic Streptomycin: 7 resistant and 12 sensitive, Izoniasin 14 resistant and 5 sensitive, rifampicin: 17 resistance and 2 sensitive and resistant to the antibiotic Ethambutol 8 and 11 sensitive. From the electrophoresis test in getting DNA band with 300bp size so that it can be concluded that 19 samples contained entirely rpoB gene.