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Molecular Characterization of Clinical Isolates to Study rpoB Gene in Mycobacterium tuberculosis Complex
One of the major health problems that can be seen around the world is Anti-microbial resistance and the main cause behind it, is easy availability and higher consumption of medicines. Rifampicin Resistance Tuberculosis (RRTB) occur when rpoB (RNA polymerase) gene in Mycobacterium tuberculosis (MTB) gets mutated and this rpoB gene encodes for beta-subunit of RNA polymerase. Clinical specimens including pulmonary and extra-pulmonary secretion were collected from patients and detected for Tuberculosis (TB) positive cases by targeting mpb64 (Mannose-Binding Protein) gene at 240 base pair. TB positive samples were taken to pre-amplification area for master-mix preparation for
Polymerase Chain Reaction (PCR) by targeting specific gene (rpoB) by using forward (5Ã¢ÂÂ-CCGCGATCAAGGAGTTCTTC-3Ã¢ÂÂ) and reverse primer (3Ã¢ÂÂ-ACACGATCTCGTCGCTAACC-5Ã¢ÂÂ). Samples were then taken to thermal cycler PCR for amplification of rpoB gene and then amplicons were taken for electrophoresis following the analyzing of amplicons in Electronic UV trans-illuminator system, in which rpoB gene was targeted at 315 base pair to determine if sample is positive for Rifampicin resistance. Author(s): Shilpa Rana, Yusra Ahmad, Narotam Sharma*, Vijay Kumar and Satish Chandra Nautiyal Abstract |