ISOLATION AND CHARACTERIZATION OF MYCOBACTERIUM TUBERCLE ISOLATES FROM PULMONARY TUBERCULOSIS PATIENTS USING CONVENTIONAL AND MOLECULAR TECHNIQUES

Abstract

Tuberculosis is a major public health issue that affects all age groups of peoples all over the world which is caused by Mycobacterium tubercle bacilli. Tuberculosis remains one of the most serious infectious disease worldwide. Dissemination of tuberculosis infection in the population occurs mainly by inhalation of contaminated aerosols from patients with active pulmonary disease. Rapid isolation and molecular characterization of Mycobacterium tubercle isolates are essential for accurate diagnosis, epidemiological surveillance and detection of drug resistance strains. The present study was carried out to isolate and characterize Mycobacterium tubercle isolates obtained from 150 pulmonary tuberculosis patients using conventional Microbiological, Biochemical and Molecular techniques. Sputum samples were processed by Ziehl – Neelsen staining and cultured on Lowenstein – Jensen (LJ) Media. Molecular confirmation was performed using Polymerase chain reaction, targeting the IS6110 Sequence.  Further characterization of isolates was carried out using Spoligotyping and MIRU- VNTR typing methods. Out of 150 sputum samples, 118 (78.6%) were smear positive, while 110 (73.3 %)yielded positive cultures. PCR confirmed 106 isolates as Mycobacterium tuberculosis. Molecular analysis demonstrated considerable genetic diversity among isolates. Multidrug- resistant strains were also identified through detection of mutations in resistance – associated genes. This study clearly demonstrates that molecular techniques are highly sensitive, rapid and reliable tools for the characterization of Mycobacterium tubercle isolates and could be significantly contribute to tuberculosis control and management programs.