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Anilkumar, A.K. and Madhavilatha , G.K. and Laiza K. , Paul and Indulakshmi, Radhakrishnan and Ajay Kumar, R. and Sathish, Mundayoor (2012) Standardization and evaluation of a tetraplex polymerase chain reaction to detect and differentiate Mycobacterium tuberculosis complex and nontuberculous Mycobacteria--a retrospective study on pulmonary TB patients. Diagnostic microbiology and infectious disease, 72 (3). pp. 239-247. ISSN 1879-0070

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Abstract

The clinical presentation of pulmonary tuberculosis by members of Mycobacterium tuberculosis complex (MTC) and nontuberculous mycobacteria (NTM) cannot be differentiated using the available standard diagnostic procedures. A single-tube tetraplex polymerase chain reaction (T-PCR) was designed to simultaneously amplify 4 well-known DNA targets of MTC. Taguchi's protocol was followed for the optimization of the conditions and was then tested on 288 pulmonary TB patient samples. The analytical sensitivity of the T-PCR was 100 fg of purified mycobacterial DNA, and specificity was found to be 100% in being able to distinguish MTC and NTM in all the cases tested. The results correlated well when validated with hsp65 PCR restriction analysis and sequencing of the 16S-23S internal transcribed spacer region, hsp65, and rpoB. The T-PCR described here is a quick, valuable, and cost-effective tool for determining whether the causative organism is MTC or NTM, and thus is useful for disease surveillance.

Item Type: Article
Uncontrolled Keywords: Mycobacterium tuberculosis complex; NTM; Multiplex PCR; Taguchi's protocol; Atypical Mycobacteria; Disease surveillance
Subjects: Mycobacterium Research
Depositing User: Rgcb Library
Date Deposited: 14 Feb 2017 06:53
Last Modified: 04 Jul 2019 07:01
URI: http://rgcb.sciencecentral.in/id/eprint/235

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