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Clinical Evaluation of Mycobacterium Tuberculosis Culture and Multiplex PCR for the Characterization of Multiple Drug Resistance Tuberculosis in Dehradun, Uttarakhand, India

Aditi Sharma, Narotam Sharma

Abstract


Multiple drug resistance (MDR) tuberculosis timely diagnose is of utmost clinical relevance and needs to be diagnose at initial stages for the proper treatment. The current study was done to detect the multiple genes for MDR tuberculosis (TB) in clinical isolates by molecular tools. 186 clinical samples were considered out and subjected for AFB smear preparation, Nested PCR (IS6110) for Mycobacterium tuberculosis complex detection and MDR TB PCR targeting rpoB, kat G, mab A promoter. Twenty-six came positive for AFB smears, out of which 08 were pulmonary and 04 were extra pulmonary. Nested PCR targeting IS6110 gene was amplified at 123 base pairs with 340 base pairs as IC (internal control) was seen in 43 cases which include 38 extra pulmonary. The Positive TB PCR specimens were subjected for MDRTB PCR Only 05 cases yielded, an amplicon of 315 bp confirming the rpoB gene resistance for resistance for rifampcin drug. In any of the 05 positives none of the other resistance gene other than rpoB was amplified. Targeting IS6110 gene and rpoB genes at once, additional information will be gained from a single test run that otherwise would require several times the reagents and more time to perform. Current study signifies the usage of quick, cost effective, DNA sequences based method for MULTIPLEX OF MDR TB detection where disease will be diagnosed earlier and hence treatment would be started at an early stage. Such experimental studies will be useful to prepare strategies for the management of the disease and to formulate the plans in Revised National Tuberculosis Control Program (RNTCP) in India as well as it will help in managing the global tuberculosis burden. The ease to execute the standardized protocols and methodologies to study the slow grower clinically relevant pathogens like M. tuberculosis.

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