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.
Copyright © 2012 Elsevier Inc. All rights reserved.
MeSH terms
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Genes, Bacterial
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Humans
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Molecular Sequence Data
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Multiplex Polymerase Chain Reaction / methods*
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Multiplex Polymerase Chain Reaction / standards*
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Mycobacterium tuberculosis / genetics
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Mycobacterium tuberculosis / isolation & purification*
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Nontuberculous Mycobacteria / genetics
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Nontuberculous Mycobacteria / isolation & purification*
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RNA, Ribosomal, 16S / genetics
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Retrospective Studies
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Sensitivity and Specificity
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Tuberculosis, Pulmonary / diagnosis*
Associated data
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GENBANK/FJ858752
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GENBANK/FJ858753
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GENBANK/FJ858754
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GENBANK/FJ858755
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