Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR
Autor(a) principal: | |
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Data de Publicação: | 2016 |
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=S1517-83822016000200506 |
Resumo: | Abstract The aim of this study was to standardize a diagnosis procedure to detect Mycobacterium avium subsp. paratuberculosis (Map) DNA in raw cow milk samples under field conditions. A procedure that combines both immunomagnetic separation and IS900 -PCR detection (IMS-IS1 PCR) was employed on milk samples from 265 lactating Holstein cows from Map infected and uninfected herds in Argentina. IMS-IS1 PCR results were analyzed and compared with those obtained from milk and fecal culture and serum ELISA. The extent of agreement between both tests was determined by the Kappa test. IMS-IS1 PCR showed a detection limit of 101 CFU of Map/mL of milk, when 50:50 mix of monoclonal and polyclonal antibodies were used to coat magnetic beads. All of the 118 samples from the Map uninfected herds were negative for the set of the tests. In Map infected herds, 80 out of 147 cows tested positive by milk IMS-IS1 PCR (55%), of which 2 (1.4%) were also positive by milk culture, 15 (10%) by fecal culture, and 20 (14%) by serum ELISA. Kappa statistics (95% CI) showed a slight agreement between the different tests (<0.20), and the proportions of agreement were ≤0.55. The IMS-IS1 PCR method detected Map in milk of the cows that were not positive in other techniques. This is the first report dealing with the application of IMS-IS1 PCR in the detection of Map in raw milk samples under field conditions in Argentina. |
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Brazilian Journal of Microbiology |
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Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCRMycobacterium avium subsp. paratuberculosisImmunomagnetic separationPCRCow's milkAbstract The aim of this study was to standardize a diagnosis procedure to detect Mycobacterium avium subsp. paratuberculosis (Map) DNA in raw cow milk samples under field conditions. A procedure that combines both immunomagnetic separation and IS900 -PCR detection (IMS-IS1 PCR) was employed on milk samples from 265 lactating Holstein cows from Map infected and uninfected herds in Argentina. IMS-IS1 PCR results were analyzed and compared with those obtained from milk and fecal culture and serum ELISA. The extent of agreement between both tests was determined by the Kappa test. IMS-IS1 PCR showed a detection limit of 101 CFU of Map/mL of milk, when 50:50 mix of monoclonal and polyclonal antibodies were used to coat magnetic beads. All of the 118 samples from the Map uninfected herds were negative for the set of the tests. In Map infected herds, 80 out of 147 cows tested positive by milk IMS-IS1 PCR (55%), of which 2 (1.4%) were also positive by milk culture, 15 (10%) by fecal culture, and 20 (14%) by serum ELISA. Kappa statistics (95% CI) showed a slight agreement between the different tests (<0.20), and the proportions of agreement were ≤0.55. The IMS-IS1 PCR method detected Map in milk of the cows that were not positive in other techniques. This is the first report dealing with the application of IMS-IS1 PCR in the detection of Map in raw milk samples under field conditions in Argentina.Sociedade Brasileira de Microbiologia2016-06-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822016000200506Brazilian Journal of Microbiology v.47 n.2 2016reponame:Brazilian Journal of Microbiologyinstname:Sociedade Brasileira de Microbiologia (SBM)instacron:SBM10.1016/j.bjm.2016.01.013info:eu-repo/semantics/openAccessGilardoni,Liliana RosaFernández,BárbaraMorsella,ClaudiaMendez,LauraJar,Ana MaríaPaolicchi,Fernando AlbertoMundo,Silvia Leonoreng2016-05-03T00:00:00Zoai:scielo:S1517-83822016000200506Revistahttps://www.scielo.br/j/bjm/ONGhttps://old.scielo.br/oai/scielo-oai.phpbjm@sbmicrobiologia.org.br||mbmartin@usp.br1678-44051517-8382opendoar:2016-05-03T00:00Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)false |
dc.title.none.fl_str_mv |
Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR |
title |
Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR |
spellingShingle |
Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR Gilardoni,Liliana Rosa Mycobacterium avium subsp. paratuberculosis Immunomagnetic separation PCR Cow's milk |
title_short |
Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR |
title_full |
Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR |
title_fullStr |
Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR |
title_full_unstemmed |
Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR |
title_sort |
Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR |
author |
Gilardoni,Liliana Rosa |
author_facet |
Gilardoni,Liliana Rosa Fernández,Bárbara Morsella,Claudia Mendez,Laura Jar,Ana María Paolicchi,Fernando Alberto Mundo,Silvia Leonor |
author_role |
author |
author2 |
Fernández,Bárbara Morsella,Claudia Mendez,Laura Jar,Ana María Paolicchi,Fernando Alberto Mundo,Silvia Leonor |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
Gilardoni,Liliana Rosa Fernández,Bárbara Morsella,Claudia Mendez,Laura Jar,Ana María Paolicchi,Fernando Alberto Mundo,Silvia Leonor |
dc.subject.por.fl_str_mv |
Mycobacterium avium subsp. paratuberculosis Immunomagnetic separation PCR Cow's milk |
topic |
Mycobacterium avium subsp. paratuberculosis Immunomagnetic separation PCR Cow's milk |
description |
Abstract The aim of this study was to standardize a diagnosis procedure to detect Mycobacterium avium subsp. paratuberculosis (Map) DNA in raw cow milk samples under field conditions. A procedure that combines both immunomagnetic separation and IS900 -PCR detection (IMS-IS1 PCR) was employed on milk samples from 265 lactating Holstein cows from Map infected and uninfected herds in Argentina. IMS-IS1 PCR results were analyzed and compared with those obtained from milk and fecal culture and serum ELISA. The extent of agreement between both tests was determined by the Kappa test. IMS-IS1 PCR showed a detection limit of 101 CFU of Map/mL of milk, when 50:50 mix of monoclonal and polyclonal antibodies were used to coat magnetic beads. All of the 118 samples from the Map uninfected herds were negative for the set of the tests. In Map infected herds, 80 out of 147 cows tested positive by milk IMS-IS1 PCR (55%), of which 2 (1.4%) were also positive by milk culture, 15 (10%) by fecal culture, and 20 (14%) by serum ELISA. Kappa statistics (95% CI) showed a slight agreement between the different tests (<0.20), and the proportions of agreement were ≤0.55. The IMS-IS1 PCR method detected Map in milk of the cows that were not positive in other techniques. This is the first report dealing with the application of IMS-IS1 PCR in the detection of Map in raw milk samples under field conditions in Argentina. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-06-01 |
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=S1517-83822016000200506 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822016000200506 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1016/j.bjm.2016.01.013 |
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.47 n.2 2016 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_ |
1752122208403062784 |