Detection of mycobacterium avium subsp. paratuberculosis by a direct in situ PCR method

Autores
Delgado, Fernando Oscar; Aguilar, Diana; Garbaccio, Sergio; Francinelli, Gladys Beatriz; Hernández Pando, R.; Romano, Maria Isabel
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
In situ detection of Mycobacterium avium subsp. paratuberculosis is useful for diagnosis and research of paratuberculosis. The aim of this paper was to detect this agent in formalin-fixed, paraffin-embedded tissue samples by a direct in situ PCR. The technique was performed on ileum or ileocaecal lymph node samples from 8 naturally infected cattle and 1 healthy calf, by using p89 and p92 primers for amplification of IS900 sequence. Moderate positive signal was detected in all positive samples and not in negative control, but tissues resulted were affected in many cases due to the enzymatic treatment and the high temperature exposition. Although the technique was useful for Map detection, the signal was lower than immunohistochemistry probably because of the fixation process. In one case, signal was higher, which might be due to the detection of spheroplasts. Thus, the described method should be recommended when others resulted negative or for spheroplasts detection.
Fil: Delgado, Fernando Oscar. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; Argentina
Fil: Aguilar, Diana. Instituto Nacional de Nutricion Salvador Zubiran; México
Fil: Garbaccio, Sergio. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; Argentina
Fil: Francinelli, Gladys Beatriz. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; Argentina
Fil: Hernández Pando, R.. Instituto Nacional de Nutricion Salvador Zubiran; México
Fil: Romano, Maria Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Materia
Paratuberculosis
PCR
Diagnosis
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/192193
Acceder
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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Detection of mycobacterium avium subsp. paratuberculosis by a direct in situ PCR methodDelgado, Fernando OscarAguilar, DianaGarbaccio, SergioFrancinelli, Gladys BeatrizHernández Pando, R.Romano, Maria IsabelParatuberculosisPCRDiagnosishttps://purl.org/becyt/ford/4.3https://purl.org/becyt/ford/4In situ detection of Mycobacterium avium subsp. paratuberculosis is useful for diagnosis and research of paratuberculosis. The aim of this paper was to detect this agent in formalin-fixed, paraffin-embedded tissue samples by a direct in situ PCR. The technique was performed on ileum or ileocaecal lymph node samples from 8 naturally infected cattle and 1 healthy calf, by using p89 and p92 primers for amplification of IS900 sequence. Moderate positive signal was detected in all positive samples and not in negative control, but tissues resulted were affected in many cases due to the enzymatic treatment and the high temperature exposition. Although the technique was useful for Map detection, the signal was lower than immunohistochemistry probably because of the fixation process. In one case, signal was higher, which might be due to the detection of spheroplasts. Thus, the described method should be recommended when others resulted negative or for spheroplasts detection.Fil: Delgado, Fernando Oscar. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; ArgentinaFil: Aguilar, Diana. Instituto Nacional de Nutricion Salvador Zubiran; MéxicoFil: Garbaccio, Sergio. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; ArgentinaFil: Francinelli, Gladys Beatriz. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; ArgentinaFil: Hernández Pando, R.. Instituto Nacional de Nutricion Salvador Zubiran; MéxicoFil: Romano, Maria Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaHindawi Publishing Corporation2011-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/192193Delgado, Fernando Oscar; Aguilar, Diana; Garbaccio, Sergio; Francinelli, Gladys Beatriz; Hernández Pando, R.; et al.; Detection of mycobacterium avium subsp. paratuberculosis by a direct in situ PCR method; Hindawi Publishing Corporation; Veterinary Medicine International; 2011; 5-2011; 1-52042-0048CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.hindawi.com/journals/vmi/2011/267102/info:eu-repo/semantics/altIdentifier/doi/10.4061/2011/267102info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:44:36Zoai:ri.conicet.gov.ar:11336/192193instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 09:44:36.666CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Detection of mycobacterium avium subsp. paratuberculosis by a direct in situ PCR method
title Detection of mycobacterium avium subsp. paratuberculosis by a direct in situ PCR method
spellingShingle Detection of mycobacterium avium subsp. paratuberculosis by a direct in situ PCR method
Delgado, Fernando Oscar
Paratuberculosis
PCR
Diagnosis
title_short Detection of mycobacterium avium subsp. paratuberculosis by a direct in situ PCR method
title_full Detection of mycobacterium avium subsp. paratuberculosis by a direct in situ PCR method
title_fullStr Detection of mycobacterium avium subsp. paratuberculosis by a direct in situ PCR method
title_full_unstemmed Detection of mycobacterium avium subsp. paratuberculosis by a direct in situ PCR method
title_sort Detection of mycobacterium avium subsp. paratuberculosis by a direct in situ PCR method
dc.creator.none.fl_str_mv Delgado, Fernando Oscar
Aguilar, Diana
Garbaccio, Sergio
Francinelli, Gladys Beatriz
Hernández Pando, R.
Romano, Maria Isabel
author Delgado, Fernando Oscar
author_facet Delgado, Fernando Oscar
Aguilar, Diana
Garbaccio, Sergio
Francinelli, Gladys Beatriz
Hernández Pando, R.
Romano, Maria Isabel
author_role author
author2 Aguilar, Diana
Garbaccio, Sergio
Francinelli, Gladys Beatriz
Hernández Pando, R.
Romano, Maria Isabel
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Paratuberculosis
PCR
Diagnosis
topic Paratuberculosis
PCR
Diagnosis
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.3
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv In situ detection of Mycobacterium avium subsp. paratuberculosis is useful for diagnosis and research of paratuberculosis. The aim of this paper was to detect this agent in formalin-fixed, paraffin-embedded tissue samples by a direct in situ PCR. The technique was performed on ileum or ileocaecal lymph node samples from 8 naturally infected cattle and 1 healthy calf, by using p89 and p92 primers for amplification of IS900 sequence. Moderate positive signal was detected in all positive samples and not in negative control, but tissues resulted were affected in many cases due to the enzymatic treatment and the high temperature exposition. Although the technique was useful for Map detection, the signal was lower than immunohistochemistry probably because of the fixation process. In one case, signal was higher, which might be due to the detection of spheroplasts. Thus, the described method should be recommended when others resulted negative or for spheroplasts detection.
Fil: Delgado, Fernando Oscar. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; Argentina
Fil: Aguilar, Diana. Instituto Nacional de Nutricion Salvador Zubiran; México
Fil: Garbaccio, Sergio. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; Argentina
Fil: Francinelli, Gladys Beatriz. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; Argentina
Fil: Hernández Pando, R.. Instituto Nacional de Nutricion Salvador Zubiran; México
Fil: Romano, Maria Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
description In situ detection of Mycobacterium avium subsp. paratuberculosis is useful for diagnosis and research of paratuberculosis. The aim of this paper was to detect this agent in formalin-fixed, paraffin-embedded tissue samples by a direct in situ PCR. The technique was performed on ileum or ileocaecal lymph node samples from 8 naturally infected cattle and 1 healthy calf, by using p89 and p92 primers for amplification of IS900 sequence. Moderate positive signal was detected in all positive samples and not in negative control, but tissues resulted were affected in many cases due to the enzymatic treatment and the high temperature exposition. Although the technique was useful for Map detection, the signal was lower than immunohistochemistry probably because of the fixation process. In one case, signal was higher, which might be due to the detection of spheroplasts. Thus, the described method should be recommended when others resulted negative or for spheroplasts detection.
publishDate 2011
dc.date.none.fl_str_mv 2011-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/192193
Delgado, Fernando Oscar; Aguilar, Diana; Garbaccio, Sergio; Francinelli, Gladys Beatriz; Hernández Pando, R.; et al.; Detection of mycobacterium avium subsp. paratuberculosis by a direct in situ PCR method; Hindawi Publishing Corporation; Veterinary Medicine International; 2011; 5-2011; 1-5
2042-0048
CONICET Digital
CONICET
url http://hdl.handle.net/11336/192193
identifier_str_mv Delgado, Fernando Oscar; Aguilar, Diana; Garbaccio, Sergio; Francinelli, Gladys Beatriz; Hernández Pando, R.; et al.; Detection of mycobacterium avium subsp. paratuberculosis by a direct in situ PCR method; Hindawi Publishing Corporation; Veterinary Medicine International; 2011; 5-2011; 1-5
2042-0048
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.hindawi.com/journals/vmi/2011/267102/
info:eu-repo/semantics/altIdentifier/doi/10.4061/2011/267102
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Hindawi Publishing Corporation
publisher.none.fl_str_mv Hindawi Publishing Corporation
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1842268677576065024
score 13.13397

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