Cloning, expression and immunological evaluation of the Omp31 protein of Brucella melitensis and evaluation of its possible use for the diagnosis in bovine brucellosis

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The serological diagnosis of brucellosis is carried out by the detection of antibodies generated against the liposaccharide «LPS» or bacterial extracts of whole cells by ELISA or agglutination tests. The objective of this study was to evaluate a recombinant protein that can be used in the serologica...

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Detalles Bibliográficos
Autores: Rosales Rangel, Jose David, Castillo Ochoa, Annie, Reyna Bello, Armando, Serrano, Ana Teresa, Fernandez Gomez, Rodolfo
Formato: artículo
Fecha de Publicación:2018
Institución:Universidad Nacional Mayor de San Marcos
Repositorio:Revistas - Universidad Nacional Mayor de San Marcos
Lenguaje:español
OAI Identifier:oai:ojs.csi.unmsm:article/14008
Enlace del recurso:https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/14008
Nivel de acceso:acceso abierto
Materia:brucellosis
Omp31r
recombinant protein
brucelosis
proteína recombinante
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spelling Cloning, expression and immunological evaluation of the Omp31 protein of Brucella melitensis and evaluation of its possible use for the diagnosis in bovine brucellosisClonación, expresión y evaluación inmunológica de la proteína Omp31 de Brucella melitensis y evaluación de su posible uso para el diagnóstico en brucelosis bovinaRosales Rangel, Jose DavidCastillo Ochoa, AnnieReyna Bello, ArmandoSerrano, Ana TeresaFernandez Gomez, RodolfobrucellosisOmp31rrecombinant proteinbrucelosisOmp31rproteína recombinanteThe serological diagnosis of brucellosis is carried out by the detection of antibodies generated against the liposaccharide «LPS» or bacterial extracts of whole cells by ELISA or agglutination tests. The objective of this study was to evaluate a recombinant protein that can be used in the serological diagnosis of bovine brucellosis. The Omp31 gene of Brucella melitensis 16M was chosen. The gene was cloned from DNA of B. melitensis; the expression of the Omp31r protein was performed in a prokaryotic expression system using the pET28a vector and purification by metal affinity chromatography (IMAC). An indirect ELISA was standardized using control sera from positive and negative bovines. The Omp31r protein reacted with the positive control sera and succeeded in discriminating against the negative sera. The sensitivity and specificity of the ELISA assay was 77.2% and 90.6%, respectively. This indirect ELISA system can be useful for the rapid diagnosis of large numbers of samples in the diagnosis of bovine brucellosis.El diagnóstico serológico de la brucelosis se lleva a cabo por la detección de anticuerpos generados contra el liposacárido “LPS” o extractos bacterianos de células enteras por ELISA o pruebas de aglutinación. El objetivo del trabajo fue evaluar una proteína recombinante que pueda ser usada en el diagnóstico serológico de la brucelosis bovina. Se escogió el gen de la Omp31 de Brucella melitensis 16M. El gen fue clonado a partir de ADN de B. melitensis. La expresión de la proteína Omp31r se realizó en un sistema de expresión procariota utilizando el vector pET28a y la purificación por cromatografía de afinidad a metales (IMAC). Un ELISA indirecto fue estandarizado utilizando sueros controles de bovinos positivos y negativos. La proteína Omp31r reaccionó con los sueros controles positivos y logró discriminarlos frente a los sueros negativos. La sensibilidad y especificidad del ensayo de ELISA fue de 77.2% y de 90.6%, respectivamente. Este sistema de ELISA indirecto puede ser útil para los diagnósticos rápidos de grandes números de muestras en el diagnóstico de la brucelosis bovina.Universidad Nacional Mayor de San Marcos, Facultad de Medicina Veterinaria2018-09-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdftext/htmlhttps://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/1400810.15381/rivep.v29i3.14008Revista de Investigaciones Veterinarias del Perú; Vol. 29 Núm. 3 (2018); 996-1008Revista de Investigaciones Veterinarias del Perú; Vol. 29 No. 3 (2018); 996-10081682-34191609-9117reponame:Revistas - Universidad Nacional Mayor de San Marcosinstname:Universidad Nacional Mayor de San Marcosinstacron:UNMSMspahttps://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/14008/13098https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/14008/13909Derechos de autor 2018 Jose David Rosales Rangelhttps://creativecommons.org/licenses/by-nc-sa/4.0info:eu-repo/semantics/openAccessoai:ojs.csi.unmsm:article/140082018-12-21T09:58:01Z
dc.title.none.fl_str_mv Cloning, expression and immunological evaluation of the Omp31 protein of Brucella melitensis and evaluation of its possible use for the diagnosis in bovine brucellosis
Clonación, expresión y evaluación inmunológica de la proteína Omp31 de Brucella melitensis y evaluación de su posible uso para el diagnóstico en brucelosis bovina
title Cloning, expression and immunological evaluation of the Omp31 protein of Brucella melitensis and evaluation of its possible use for the diagnosis in bovine brucellosis
spellingShingle Cloning, expression and immunological evaluation of the Omp31 protein of Brucella melitensis and evaluation of its possible use for the diagnosis in bovine brucellosis
Rosales Rangel, Jose David
brucellosis
Omp31r
recombinant protein
brucelosis
Omp31r
proteína recombinante
title_short Cloning, expression and immunological evaluation of the Omp31 protein of Brucella melitensis and evaluation of its possible use for the diagnosis in bovine brucellosis
title_full Cloning, expression and immunological evaluation of the Omp31 protein of Brucella melitensis and evaluation of its possible use for the diagnosis in bovine brucellosis
title_fullStr Cloning, expression and immunological evaluation of the Omp31 protein of Brucella melitensis and evaluation of its possible use for the diagnosis in bovine brucellosis
title_full_unstemmed Cloning, expression and immunological evaluation of the Omp31 protein of Brucella melitensis and evaluation of its possible use for the diagnosis in bovine brucellosis
title_sort Cloning, expression and immunological evaluation of the Omp31 protein of Brucella melitensis and evaluation of its possible use for the diagnosis in bovine brucellosis
dc.creator.none.fl_str_mv Rosales Rangel, Jose David
Castillo Ochoa, Annie
Reyna Bello, Armando
Serrano, Ana Teresa
Fernandez Gomez, Rodolfo
author Rosales Rangel, Jose David
author_facet Rosales Rangel, Jose David
Castillo Ochoa, Annie
Reyna Bello, Armando
Serrano, Ana Teresa
Fernandez Gomez, Rodolfo
author_role author
author2 Castillo Ochoa, Annie
Reyna Bello, Armando
Serrano, Ana Teresa
Fernandez Gomez, Rodolfo
author2_role author
author
author
author
dc.subject.none.fl_str_mv brucellosis
Omp31r
recombinant protein
brucelosis
Omp31r
proteína recombinante
topic brucellosis
Omp31r
recombinant protein
brucelosis
Omp31r
proteína recombinante
description The serological diagnosis of brucellosis is carried out by the detection of antibodies generated against the liposaccharide «LPS» or bacterial extracts of whole cells by ELISA or agglutination tests. The objective of this study was to evaluate a recombinant protein that can be used in the serological diagnosis of bovine brucellosis. The Omp31 gene of Brucella melitensis 16M was chosen. The gene was cloned from DNA of B. melitensis; the expression of the Omp31r protein was performed in a prokaryotic expression system using the pET28a vector and purification by metal affinity chromatography (IMAC). An indirect ELISA was standardized using control sera from positive and negative bovines. The Omp31r protein reacted with the positive control sera and succeeded in discriminating against the negative sera. The sensitivity and specificity of the ELISA assay was 77.2% and 90.6%, respectively. This indirect ELISA system can be useful for the rapid diagnosis of large numbers of samples in the diagnosis of bovine brucellosis.
publishDate 2018
dc.date.none.fl_str_mv 2018-09-06
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/14008
10.15381/rivep.v29i3.14008
url https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/14008
identifier_str_mv 10.15381/rivep.v29i3.14008
dc.language.none.fl_str_mv spa
language spa
dc.relation.none.fl_str_mv https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/14008/13098
https://revistasinvestigacion.unmsm.edu.pe/index.php/veterinaria/article/view/14008/13909
dc.rights.none.fl_str_mv Derechos de autor 2018 Jose David Rosales Rangel
https://creativecommons.org/licenses/by-nc-sa/4.0
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Derechos de autor 2018 Jose David Rosales Rangel
https://creativecommons.org/licenses/by-nc-sa/4.0
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
text/html
dc.publisher.none.fl_str_mv Universidad Nacional Mayor de San Marcos, Facultad de Medicina Veterinaria
publisher.none.fl_str_mv Universidad Nacional Mayor de San Marcos, Facultad de Medicina Veterinaria
dc.source.none.fl_str_mv Revista de Investigaciones Veterinarias del Perú; Vol. 29 Núm. 3 (2018); 996-1008
Revista de Investigaciones Veterinarias del Perú; Vol. 29 No. 3 (2018); 996-1008
1682-3419
1609-9117
reponame:Revistas - Universidad Nacional Mayor de San Marcos
instname:Universidad Nacional Mayor de San Marcos
instacron:UNMSM
instname_str Universidad Nacional Mayor de San Marcos
instacron_str UNMSM
institution UNMSM
reponame_str Revistas - Universidad Nacional Mayor de San Marcos
collection Revistas - Universidad Nacional Mayor de San Marcos
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 13.982926
Cloning, expression and immunological evaluation of the Omp31 protein of Brucella melitensis and evaluation of its possible use for the diagnosis in bovine brucellosis
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