A novel multiplex real-time PCR for the identification of mycobacteria associated with zoonotic tuberculosis.

<h4>Background</h4>Tuberculosis (TB) is the leading cause of death worldwide from a single infectious agent. An ability to detect the Mycobacterium tuberculosis complex (MTC) in clinical material while simultaneously differentiating its members is considered important. This allows for th...

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Autores principales:	Kate Reddington, Justin O'Grady, Siobhan Dorai-Raj, Stefan Niemann, Dick van Soolingen, Thomas Barry
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c42412d464c44eab95b5a882d42ce2b0
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spelling	oai:doaj.org-article:c42412d464c44eab95b5a882d42ce2b02021-11-18T06:48:23ZA novel multiplex real-time PCR for the identification of mycobacteria associated with zoonotic tuberculosis.1932-620310.1371/journal.pone.0023481https://doaj.org/article/c42412d464c44eab95b5a882d42ce2b02011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21858140/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Tuberculosis (TB) is the leading cause of death worldwide from a single infectious agent. An ability to detect the Mycobacterium tuberculosis complex (MTC) in clinical material while simultaneously differentiating its members is considered important. This allows for the gathering of epidemiological information pertaining to the prevalence, transmission and geographical distribution of the MTC, including those MTC members associated with zoonotic TB infection in humans. Also differentiating between members of the MTC provides the clinician with inherent MTC specific drug susceptibility profiles to guide appropriate chemotherapy.<h4>Methodology/principal findings</h4>The aim of this study was to develop a multiplex real-time PCR assay using novel molecular targets to identify and differentiate between the phylogenetically closely related M. bovis, M. bovis BCG and M. caprae. The lpqT gene was explored for the collective identification of M. bovis, M. bovis BCG and M. caprae, the lepA gene was targeted for the specific identification of M. caprae and a Region of Difference 1 (RD1) assay was incorporated in the test to differentiate M. bovis BCG. The multiplex real-time PCR assay was evaluated on 133 bacterial strains and was determined to be 100% specific for the members of the MTC targeted.<h4>Conclusions/significance</h4>The multiplex real-time PCR assay developed in this study is the first assay described for the identification and simultaneous differentiation of M. bovis, M. bovis BCG and M. caprae in one internally controlled reaction. Future validation of this multiplex assay should demonstrate its potential in the rapid and accurate diagnosis of TB caused by these three mycobacteria. Furthermore, the developed assay may be used in conjunction with a recently described multiplex real-time PCR assay for identification of the MTC and simultaneous differentiation of M. tuberculosis, M. canettii resulting in an ability to differentiate five of the eight members of the MTC.Kate ReddingtonJustin O'GradySiobhan Dorai-RajStefan NiemannDick van SoolingenThomas BarryPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 8, p e23481 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Kate Reddington Justin O'Grady Siobhan Dorai-Raj Stefan Niemann Dick van Soolingen Thomas Barry A novel multiplex real-time PCR for the identification of mycobacteria associated with zoonotic tuberculosis.
description	<h4>Background</h4>Tuberculosis (TB) is the leading cause of death worldwide from a single infectious agent. An ability to detect the Mycobacterium tuberculosis complex (MTC) in clinical material while simultaneously differentiating its members is considered important. This allows for the gathering of epidemiological information pertaining to the prevalence, transmission and geographical distribution of the MTC, including those MTC members associated with zoonotic TB infection in humans. Also differentiating between members of the MTC provides the clinician with inherent MTC specific drug susceptibility profiles to guide appropriate chemotherapy.<h4>Methodology/principal findings</h4>The aim of this study was to develop a multiplex real-time PCR assay using novel molecular targets to identify and differentiate between the phylogenetically closely related M. bovis, M. bovis BCG and M. caprae. The lpqT gene was explored for the collective identification of M. bovis, M. bovis BCG and M. caprae, the lepA gene was targeted for the specific identification of M. caprae and a Region of Difference 1 (RD1) assay was incorporated in the test to differentiate M. bovis BCG. The multiplex real-time PCR assay was evaluated on 133 bacterial strains and was determined to be 100% specific for the members of the MTC targeted.<h4>Conclusions/significance</h4>The multiplex real-time PCR assay developed in this study is the first assay described for the identification and simultaneous differentiation of M. bovis, M. bovis BCG and M. caprae in one internally controlled reaction. Future validation of this multiplex assay should demonstrate its potential in the rapid and accurate diagnosis of TB caused by these three mycobacteria. Furthermore, the developed assay may be used in conjunction with a recently described multiplex real-time PCR assay for identification of the MTC and simultaneous differentiation of M. tuberculosis, M. canettii resulting in an ability to differentiate five of the eight members of the MTC.
format	article
author	Kate Reddington Justin O'Grady Siobhan Dorai-Raj Stefan Niemann Dick van Soolingen Thomas Barry
author_facet	Kate Reddington Justin O'Grady Siobhan Dorai-Raj Stefan Niemann Dick van Soolingen Thomas Barry
author_sort	Kate Reddington
title	A novel multiplex real-time PCR for the identification of mycobacteria associated with zoonotic tuberculosis.
title_short	A novel multiplex real-time PCR for the identification of mycobacteria associated with zoonotic tuberculosis.
title_full	A novel multiplex real-time PCR for the identification of mycobacteria associated with zoonotic tuberculosis.
title_fullStr	A novel multiplex real-time PCR for the identification of mycobacteria associated with zoonotic tuberculosis.
title_full_unstemmed	A novel multiplex real-time PCR for the identification of mycobacteria associated with zoonotic tuberculosis.
title_sort	novel multiplex real-time pcr for the identification of mycobacteria associated with zoonotic tuberculosis.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/c42412d464c44eab95b5a882d42ce2b0
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A novel multiplex real-time PCR for the identification of mycobacteria associated with zoonotic tuberculosis.

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