Cargando…
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...
Autores principales: | , , , , , , , , |
---|---|
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 |
_version_ | 1782400962894233600 |
---|---|
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. |
format | Online Article Text |
id | pubmed-4646627 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT housmangenevieve validationofqpcrmethodsforthedetectionofmycobacteriuminnewworldanimalreservoirs AT malukiewiczjoanna validationofqpcrmethodsforthedetectionofmycobacteriuminnewworldanimalreservoirs AT boerevanner validationofqpcrmethodsforthedetectionofmycobacteriuminnewworldanimalreservoirs AT grativoladrianad validationofqpcrmethodsforthedetectionofmycobacteriuminnewworldanimalreservoirs AT pereiraluizcezarm validationofqpcrmethodsforthedetectionofmycobacteriuminnewworldanimalreservoirs AT silvaitadeoliveirae validationofqpcrmethodsforthedetectionofmycobacteriuminnewworldanimalreservoirs AT ruizmirandacarlosr validationofqpcrmethodsforthedetectionofmycobacteriuminnewworldanimalreservoirs AT trumanrichard validationofqpcrmethodsforthedetectionofmycobacteriuminnewworldanimalreservoirs AT stoneannec validationofqpcrmethodsforthedetectionofmycobacteriuminnewworldanimalreservoirs |