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Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs

Zoonotic pathogens that cause leprosy (Mycobacterium leprae) and tuberculosis (Mycobacterium tuberculosis complex, MTBC) continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human expos...

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Autores principales: Housman, Genevieve, Malukiewicz, Joanna, Boere, Vanner, Grativol, Adriana D., Pereira, Luiz Cezar M., Silva, Ita de Oliveira e, Ruiz-Miranda, Carlos R., Truman, Richard, Stone, Anne C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4646627/
https://www.ncbi.nlm.nih.gov/pubmed/26571269
http://dx.doi.org/10.1371/journal.pntd.0004198
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author Housman, Genevieve
Malukiewicz, Joanna
Boere, Vanner
Grativol, Adriana D.
Pereira, Luiz Cezar M.
Silva, Ita de Oliveira e
Ruiz-Miranda, Carlos R.
Truman, Richard
Stone, Anne C.
author_facet Housman, Genevieve
Malukiewicz, Joanna
Boere, Vanner
Grativol, Adriana D.
Pereira, Luiz Cezar M.
Silva, Ita de Oliveira e
Ruiz-Miranda, Carlos R.
Truman, Richard
Stone, Anne C.
author_sort Housman, Genevieve
collection PubMed
description Zoonotic pathogens that cause leprosy (Mycobacterium leprae) and tuberculosis (Mycobacterium tuberculosis complex, MTBC) continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR. In a trial study in which armadillos were artificially infected with M. leprae, these techniques were specific and sensitive to pathogen detection, while more traditional ELISAs were only specific. These assays were then employed in a case study to detect M. leprae as well as MTBC in wild marmosets. All marmosets were negative for M. leprae DNA, but 14 were positive for the mycobacterial rpoB gene assay. Targeted capture and sequencing of rpoB and other MTBC genes validated the presence of mycobacterial DNA in these samples and revealed that qPCR is useful for identifying mycobacterial-infected animal hosts.
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spelling pubmed-46466272015-11-25 Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs Housman, Genevieve Malukiewicz, Joanna Boere, Vanner Grativol, Adriana D. Pereira, Luiz Cezar M. Silva, Ita de Oliveira e Ruiz-Miranda, Carlos R. Truman, Richard Stone, Anne C. PLoS Negl Trop Dis Research Article Zoonotic pathogens that cause leprosy (Mycobacterium leprae) and tuberculosis (Mycobacterium tuberculosis complex, MTBC) continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR. In a trial study in which armadillos were artificially infected with M. leprae, these techniques were specific and sensitive to pathogen detection, while more traditional ELISAs were only specific. These assays were then employed in a case study to detect M. leprae as well as MTBC in wild marmosets. All marmosets were negative for M. leprae DNA, but 14 were positive for the mycobacterial rpoB gene assay. Targeted capture and sequencing of rpoB and other MTBC genes validated the presence of mycobacterial DNA in these samples and revealed that qPCR is useful for identifying mycobacterial-infected animal hosts. Public Library of Science 2015-11-16 /pmc/articles/PMC4646627/ /pubmed/26571269 http://dx.doi.org/10.1371/journal.pntd.0004198 Text en https://creativecommons.org/publicdomain/zero/1.0/ This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.
spellingShingle Research Article
Housman, Genevieve
Malukiewicz, Joanna
Boere, Vanner
Grativol, Adriana D.
Pereira, Luiz Cezar M.
Silva, Ita de Oliveira e
Ruiz-Miranda, Carlos R.
Truman, Richard
Stone, Anne C.
Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs
title Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs
title_full Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs
title_fullStr Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs
title_full_unstemmed Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs
title_short Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs
title_sort validation of qpcr methods for the detection of mycobacterium in new world animal reservoirs
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4646627/
https://www.ncbi.nlm.nih.gov/pubmed/26571269
http://dx.doi.org/10.1371/journal.pntd.0004198
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