Tuberculosis is a matter of concern for all countries. Tuberculosis (TB) remains one of the major global health threats leading to morbidity and mortality. One in three persons across the world representing 2–3 billion individuals are known to be infected with Mycobacterium Tuberculosis (M. Tuberculosis) of which 5–15% are likely to develop active TB disease during their lifetime.Information of the pattern of drug resistant among the tuberculosis is crucial in developing countries. Therefore, this study aims to assess the drug resistance pattern of Mycobacterium tuberculosis isolates and associated factors among the patients. This study selected 934 culturepositive sputum samples referred to the National Reference Laboratory of the Research Institute for Pulmonology in Thanjavur from January 2015 to January 2018 were analysed; 40% of these samples were obtained from Tb Sanatorium, Sengipattipatients (hospitalized in `Sengipatti) and 60% from other regions (hospitalized in Minsk and other regions) equal to patient’s population in the regions.All 934 cases were subjected to a drug-resistance test. The anti-microbial drug susceptibility tests (DST) were performed using the WHO standard conventional proportional method. The Preferable First Line Drugs were INH 1mcg/ml, RIF40 mcg/ml, Ethambutol (EMB) 2 mcg/ml, and Streptomycin (SM) 10 mcg/ml on slants with the H37Rv strain of MTB as the positive control. Furthermore, MDR isolates were tested for resistance to fluoroquinolones and three injectable drugs (Amikacin 8 mcg/ml, Kanamycin 30 mcg/ml, and Capreomycin 8mcg/ml) for detection of XDR isolates.