RESEARCH ARTICLE


Rapid Identification of Mycobacterium Species with the Aid of Multiplex Polymerase Chain Reaction (PCR) From Clinical Isolates



Siddhartha Gupta1, Debasis Bandyopadhyay1, *, Suman Kalyan Paine1, Soma Gupta2, Surajita Banerjee1, Sujata Bhattacharya3, Ratan Gachhui4, Basudev Bhattacharya1
1 Biochemistry Research Wing, Department of Biochemistry, IPGME & R, SSKM Hospital, Kolkata, India
2 Department of Biochemistry, Midnapore Medical College, Midnapore, India
3 Department of Microbiology, R G Kar Medical College, Kolkata, India
4 Department of Life Science & Biotechnology, Jadavpur University, India


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© Gupta et al.; Licensee Bentham Open.

open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

* Address correspondence to this author at the Biochemistry Research Wing, Department of Biochemistry, IPGME & R, SSKM Hospital, Kolkata, India; Tel: +913322236717; E-mail: debasis_94@rocketmail.com


Abstract

Mycobacteria are aerobic, nonspore forming, non-motile,single-cell bacteria.Of more than 40 currently recognized species of mycobacteria, Mycobacterium tuberculosis, the causative agent of human TB is the commonest pathogen for pulmonary and extra pulmonary tuberculosis cases. The other members of the Mycobacterium tuberculosis complex (MTC) or the nontubercular mycobacterium (NTM) produces similar diseases which cannot be differentiated from tuberculosis by clinical symptoms and signs. But this differentiation is important as the chemotherapy varies widely according to the strain of mycobacterium. The burden of morbidity and mortality of tuberculosis is rapidly growing worldwide, particularly with the HIV/AIDS epidemic. The strain identification of Mycobacterium remains a cumbersome, labor intensive and expensive procedure, which requires 3 to 12 weeks of time. The conventional methods of strain identification lack proper standardization and precise diagnosis. The prime objective of this study is to overcome these problems.

A multiplex PCR using 3 amplicons of 165,365, and 541 base pair target sequences was done with a total number of 165 clinical isolates of suspected Koch’s patients. Strain identification was compared both by conventional methods and multiplex PCR. The results of the study show that this multiplex PCR is supposed to be less complicated, less time consuming, cost-effective and superior to the conventional methods. It is also applicable for culture negative samples where strain identification is not possible by conventional approach.

Key Words: M. tuberculosis, Non Tubercular Mycobacteriosis, Culture, Multiplex PCR, Strain differention.