Abstract

The ineffectiveness of drugs like isoniazid and rifampicin which are first line of drugs targeted at the treatment of Mycobacterium tuberculosis has increased over the years despite decrease in cases of Mycobacterium tuberculosis either due to the patients not following the right course of treatment or increase in adaptation of the pathogens to the anti-tuberculosis drugs. The aim of this study was the use of Molecular and Microbiological Assays for Characterization of Multi-Drug Resistant Tuberculosis by Multiplex Polymerase Chain Reaction method - Clinical implementation for the Disease Diagnosis and Treatment. 62 clinical samples were collected from symptomatic patients and the processes of decontamination of samples using modified Petroffe method, Ziehl– Neelsen stains (Acid Fast Bacilli Stain, AFB) for initial detection, culturing of clinical samples in Lowenstein-Jensen media, DNA isolation using solid phase (silica column) and magnetic beads methods, Master Mix preparation and PCR set up for multiplex PCR, detection of amplicons using post amplification method and E-gel imager or gel documentation system were followed accordingly. The results of this study showed that out of 62 clinical samples processed, 26(41.93%) stained positive in Acid Fast Bacilli Stain, visible growths were observed on the cultures after 5 weeks and 13(20.97%) came positive in Multiplex Polymerase Chain Reaction (MPCR). Though the initial molecular detection of Mycobacterium tuberculosis is done targeting IS6110 and mpb6 in Nested PCR method, these have never been specific test for detection of drug resistant genes in Mycobacterium tuberculosis Complex (MTC) and therefore ineffective line of drugs may be administered allowing the disease to persist. The use of MPCR is a specific and better way for the diagnosis as well as treatment of Mycobacterium tuberculosis.

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