LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis
Autor(a) principal: | |
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Data de Publicação: | 2015 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Brazilian Journal of Microbiology |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822015000200619 |
Resumo: | In this study, we developed new sets of primers to detect Brucella spp. and M. avium subsp. paratuberculosis (MAP) through isothermal amplification. We selected a previously well-characterized target gene, bscp31, specific for Brucella spp. and IS900 for MAP. The limits of detection using the loop-mediated isothermal amplification (LAMP) protocols described herein were similar to those of conventional PCR targeting the same sequences. Hydroxynaphtol blue and SYBR GreenTM allowed direct naked-eye detection with identical sensitivity as agarose gel electrophoresis. We included the LAMP-based protocol in a rapid identification scheme of the respective pathogens, and all tested isolates were correctly identified within 2 to 3 h. In addition, both protocols were suitable for specifically identifying the respective pathogens; in the case of Brucella, it also allowed the identification of all the biovars tested. We conclude that LAMP is a suitable rapid molecular typing tool that could help to shorten the time required to identify insidious bacteria in low-complexity laboratories, mainly in developing countries. |
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Brazilian Journal of Microbiology |
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LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosisloop-mediated isothermal amplificationmolecular typingbrucellosisparatuberculosisIn this study, we developed new sets of primers to detect Brucella spp. and M. avium subsp. paratuberculosis (MAP) through isothermal amplification. We selected a previously well-characterized target gene, bscp31, specific for Brucella spp. and IS900 for MAP. The limits of detection using the loop-mediated isothermal amplification (LAMP) protocols described herein were similar to those of conventional PCR targeting the same sequences. Hydroxynaphtol blue and SYBR GreenTM allowed direct naked-eye detection with identical sensitivity as agarose gel electrophoresis. We included the LAMP-based protocol in a rapid identification scheme of the respective pathogens, and all tested isolates were correctly identified within 2 to 3 h. In addition, both protocols were suitable for specifically identifying the respective pathogens; in the case of Brucella, it also allowed the identification of all the biovars tested. We conclude that LAMP is a suitable rapid molecular typing tool that could help to shorten the time required to identify insidious bacteria in low-complexity laboratories, mainly in developing countries.Sociedade Brasileira de Microbiologia2015-06-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822015000200619Brazilian Journal of Microbiology v.46 n.2 2015reponame:Brazilian Journal of Microbiologyinstname:Sociedade Brasileira de Microbiologia (SBM)instacron:SBM10.1590/S1517-838246220131206info:eu-repo/semantics/openAccessTrangoni,Marcos D.Gioffré,Andrea K.Cerón Cucchi,María E.Caimi,Karina C.Ruybal,PaulaZumárraga,Martín J.Cravero,Silvio L.eng2015-11-27T00:00:00Zoai:scielo:S1517-83822015000200619Revistahttps://www.scielo.br/j/bjm/ONGhttps://old.scielo.br/oai/scielo-oai.phpbjm@sbmicrobiologia.org.br||mbmartin@usp.br1678-44051517-8382opendoar:2015-11-27T00:00Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)false |
dc.title.none.fl_str_mv |
LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
title |
LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
spellingShingle |
LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis Trangoni,Marcos D. loop-mediated isothermal amplification molecular typing brucellosis paratuberculosis |
title_short |
LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
title_full |
LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
title_fullStr |
LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
title_full_unstemmed |
LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
title_sort |
LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
author |
Trangoni,Marcos D. |
author_facet |
Trangoni,Marcos D. Gioffré,Andrea K. Cerón Cucchi,María E. Caimi,Karina C. Ruybal,Paula Zumárraga,Martín J. Cravero,Silvio L. |
author_role |
author |
author2 |
Gioffré,Andrea K. Cerón Cucchi,María E. Caimi,Karina C. Ruybal,Paula Zumárraga,Martín J. Cravero,Silvio L. |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
Trangoni,Marcos D. Gioffré,Andrea K. Cerón Cucchi,María E. Caimi,Karina C. Ruybal,Paula Zumárraga,Martín J. Cravero,Silvio L. |
dc.subject.por.fl_str_mv |
loop-mediated isothermal amplification molecular typing brucellosis paratuberculosis |
topic |
loop-mediated isothermal amplification molecular typing brucellosis paratuberculosis |
description |
In this study, we developed new sets of primers to detect Brucella spp. and M. avium subsp. paratuberculosis (MAP) through isothermal amplification. We selected a previously well-characterized target gene, bscp31, specific for Brucella spp. and IS900 for MAP. The limits of detection using the loop-mediated isothermal amplification (LAMP) protocols described herein were similar to those of conventional PCR targeting the same sequences. Hydroxynaphtol blue and SYBR GreenTM allowed direct naked-eye detection with identical sensitivity as agarose gel electrophoresis. We included the LAMP-based protocol in a rapid identification scheme of the respective pathogens, and all tested isolates were correctly identified within 2 to 3 h. In addition, both protocols were suitable for specifically identifying the respective pathogens; in the case of Brucella, it also allowed the identification of all the biovars tested. We conclude that LAMP is a suitable rapid molecular typing tool that could help to shorten the time required to identify insidious bacteria in low-complexity laboratories, mainly in developing countries. |
publishDate |
2015 |
dc.date.none.fl_str_mv |
2015-06-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822015000200619 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822015000200619 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S1517-838246220131206 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Microbiologia |
publisher.none.fl_str_mv |
Sociedade Brasileira de Microbiologia |
dc.source.none.fl_str_mv |
Brazilian Journal of Microbiology v.46 n.2 2015 reponame:Brazilian Journal of Microbiology instname:Sociedade Brasileira de Microbiologia (SBM) instacron:SBM |
instname_str |
Sociedade Brasileira de Microbiologia (SBM) |
instacron_str |
SBM |
institution |
SBM |
reponame_str |
Brazilian Journal of Microbiology |
collection |
Brazilian Journal of Microbiology |
repository.name.fl_str_mv |
Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM) |
repository.mail.fl_str_mv |
bjm@sbmicrobiologia.org.br||mbmartin@usp.br |
_version_ |
1752122207494995968 |