Aim: The aim of this study was to profile the anti-microbial resistant of Mycobacterial strains causing cluster pulmonary infections in Bayelsa State, Nigeria. Study Design: Cross sectional facility based. Place and Duration: This study was carried out in all the in tuberculosis centers in all the eight local government area in Bayelsa State, Nigeria between January 2019 to February 2021. Materials and Methods: A total of 102 tuberculosis patients who had been previously diagnosed of Tuberculosis (both new and old cases) participated in the study. Two sputum samples were collected from each subject in a wide mouth screw cape container and falcon tube container respectively. The first was used for GenXpert technique while the second was used for culture on Lowenstein Jensen solid Media. The visible and confirmed Mycobacterium growth was subjected to Line Probe Assay (MTBDRplus assay) and genetic sequencing. The DNA of the isolates where extracted using Genolyse method. The extracted DNA was used to perform a gene mutation profiling using MTBRplus Assay. The 16Sr RNA sequencing was done on the amplified genes using the BigDye terminator kit on a 3510 AB1 sequencer. Results: Out of the 102 sputum samples analyzed a total of 91(89.2%) were GeneXpert positive. Drug resistant profiling by GeneXpert shows 8(7.8) strain mutation at the rpoB gene only while the resistant profiling of the isolated on Lowenstein Jensen solid media had 14(13.7) strain with mutation on genes responsible for first and second line drug resistant. Two non-tuberculosis Mycobacterium species which are Mycobacteriode abscessus subsp. abscessus st and Mycobacterium Kansasii strain FDAARGOS_1534 where isolated among Tuberculosis patients, both are multidrug-resistant strains. The Mycobacterium tuberculosis strains in circulation causing cluster TB infection in Yenagoa especially and other parts of Bayelsa State, Nigeria are MG003 and R2092 strains, but the most predominant strain that is traced to cluster TB infection is MG003. The extraction of MTB gene from a pure culture of a Lowenstein Jensen selected media reduced the possibility of contamination and enhanced the reliability of gene sequencing and Bioinformatics analysis which includes comparing strains of MTB with the once in the National center for Biotechnology information (BLAST) data base. Conclusion: The resistance profiling of MTB reveals that most strains were resistant to rifampicin. This means that there were more mutation on the rifampicin activation gene (rpoB) than any other gene. Most cases of multi drug resistance is associated with rifampicin resistance while cases of extensive drug resistance is not common.