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Tuberculosis (TB) infections caused by multidrug-resistant Mycobacterium tuberculosis(MDR MTB) remain a significant public health concern worldwide. Georgia has a high prevalence of MDR MTB. The genetic mechanisms underlying the emergence of MDR MTB strains in this region are poorly understood and need to be determined for developing better strategies for TB control. This study investigated the frequency of major drug resistance mutations across rpoB, katGand inhAloci of Georgian MDR MTB strains and explored differences between new and previously treated patients.