Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease

Autores
Larsen, Alejandra; Corva, Santiago; Panei, Carlos Javier; Geisler, Christoph; Mortola, Eduardo
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Enzyme-linked immunosorbent assay (ELISA) is often used to test bovineleukemia virus (BLV) infection. However, commercially available kits test inSouth America detect only antibodies against the gp51 protein. With the aimto improve the sensitivity of the test, we developed here a two-step indirectdual ELISA test that included both proteins p24 and gp51, expressed andproduced in E. coli and baculovirus expression system respectively. Two hundredten BLV sera, stated as double positive or double negative by the combinationof commercial agar gel immunodiffusion (AGID) assay and a gp51-ELISA test, were tested with our in house dual rp24/rgp51 ELISA. Firstly, we checked the purified, optimized and standardized proteins as antigen by the checkerboard technique, and set up our in house ELISA test. The concordancecorrelation coefficient (CCC) and coefficient of variation (CV) intraplate repeatability levels were within the values established by the international standards.The statistical analysis demonstrated the value of sera correctly rankedhighest (93.48%), and for 0.3 cutoff, the sensitivity was 95.65% and the specificity 91.30%. In conclusion, the rp24/rgp51 ELISA developed and standardized here demonstrated to have good analytical characteristics to be considered for screening of BLV.
Fil: Larsen, Alejandra. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Fil: Corva, Santiago. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Fil: Panei, Carlos Javier. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina
Fil: Geisler, Christoph. University Of Wyoming; Estados Unidos
Fil: Mortola, Eduardo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Materia
BOVINE LEUKEMIA VIRUS,
INDIRECT DUAL ELISA
RECOMBINANT ANTIGENS
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:11336/49546

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network_name_str CONICET Digital (CONICET)
spelling Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the DiseaseLarsen, AlejandraCorva, SantiagoPanei, Carlos JavierGeisler, ChristophMortola, EduardoBOVINE LEUKEMIA VIRUS,INDIRECT DUAL ELISARECOMBINANT ANTIGENShttps://purl.org/becyt/ford/4.4https://purl.org/becyt/ford/4Enzyme-linked immunosorbent assay (ELISA) is often used to test bovineleukemia virus (BLV) infection. However, commercially available kits test inSouth America detect only antibodies against the gp51 protein. With the aimto improve the sensitivity of the test, we developed here a two-step indirectdual ELISA test that included both proteins p24 and gp51, expressed andproduced in E. coli and baculovirus expression system respectively. Two hundredten BLV sera, stated as double positive or double negative by the combinationof commercial agar gel immunodiffusion (AGID) assay and a gp51-ELISA test, were tested with our in house dual rp24/rgp51 ELISA. Firstly, we checked the purified, optimized and standardized proteins as antigen by the checkerboard technique, and set up our in house ELISA test. The concordancecorrelation coefficient (CCC) and coefficient of variation (CV) intraplate repeatability levels were within the values established by the international standards.The statistical analysis demonstrated the value of sera correctly rankedhighest (93.48%), and for 0.3 cutoff, the sensitivity was 95.65% and the specificity 91.30%. In conclusion, the rp24/rgp51 ELISA developed and standardized here demonstrated to have good analytical characteristics to be considered for screening of BLV.Fil: Larsen, Alejandra. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaFil: Corva, Santiago. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaFil: Panei, Carlos Javier. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; ArgentinaFil: Geisler, Christoph. University Of Wyoming; Estados UnidosFil: Mortola, Eduardo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaScientific Research Publishing Inc2017-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/49546Larsen, Alejandra; Corva, Santiago; Panei, Carlos Javier; Geisler, Christoph; Mortola, Eduardo; Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease; Scientific Research Publishing Inc; Open Journal of Animal Sciences; 07; 03; 7-2017; 241-2532161-75972161-7627CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.scirp.org/journal/doi.aspx?DOI=10.4236/ojas.2017.73019info:eu-repo/semantics/altIdentifier/doi/10.4236/ojas.2017.73019info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:11:20Zoai:ri.conicet.gov.ar:11336/49546instacron: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:34982025-10-15 15:11:20.76CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease
title Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease
spellingShingle Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease
Larsen, Alejandra
BOVINE LEUKEMIA VIRUS,
INDIRECT DUAL ELISA
RECOMBINANT ANTIGENS
title_short Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease
title_full Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease
title_fullStr Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease
title_full_unstemmed Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease
title_sort Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease
dc.creator.none.fl_str_mv Larsen, Alejandra
Corva, Santiago
Panei, Carlos Javier
Geisler, Christoph
Mortola, Eduardo
author Larsen, Alejandra
author_facet Larsen, Alejandra
Corva, Santiago
Panei, Carlos Javier
Geisler, Christoph
Mortola, Eduardo
author_role author
author2 Corva, Santiago
Panei, Carlos Javier
Geisler, Christoph
Mortola, Eduardo
author2_role author
author
author
author
dc.subject.none.fl_str_mv BOVINE LEUKEMIA VIRUS,
INDIRECT DUAL ELISA
RECOMBINANT ANTIGENS
topic BOVINE LEUKEMIA VIRUS,
INDIRECT DUAL ELISA
RECOMBINANT ANTIGENS
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.4
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv Enzyme-linked immunosorbent assay (ELISA) is often used to test bovineleukemia virus (BLV) infection. However, commercially available kits test inSouth America detect only antibodies against the gp51 protein. With the aimto improve the sensitivity of the test, we developed here a two-step indirectdual ELISA test that included both proteins p24 and gp51, expressed andproduced in E. coli and baculovirus expression system respectively. Two hundredten BLV sera, stated as double positive or double negative by the combinationof commercial agar gel immunodiffusion (AGID) assay and a gp51-ELISA test, were tested with our in house dual rp24/rgp51 ELISA. Firstly, we checked the purified, optimized and standardized proteins as antigen by the checkerboard technique, and set up our in house ELISA test. The concordancecorrelation coefficient (CCC) and coefficient of variation (CV) intraplate repeatability levels were within the values established by the international standards.The statistical analysis demonstrated the value of sera correctly rankedhighest (93.48%), and for 0.3 cutoff, the sensitivity was 95.65% and the specificity 91.30%. In conclusion, the rp24/rgp51 ELISA developed and standardized here demonstrated to have good analytical characteristics to be considered for screening of BLV.
Fil: Larsen, Alejandra. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Fil: Corva, Santiago. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Fil: Panei, Carlos Javier. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Microbiología. Cátedra de Virología; Argentina
Fil: Geisler, Christoph. University Of Wyoming; Estados Unidos
Fil: Mortola, Eduardo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
description Enzyme-linked immunosorbent assay (ELISA) is often used to test bovineleukemia virus (BLV) infection. However, commercially available kits test inSouth America detect only antibodies against the gp51 protein. With the aimto improve the sensitivity of the test, we developed here a two-step indirectdual ELISA test that included both proteins p24 and gp51, expressed andproduced in E. coli and baculovirus expression system respectively. Two hundredten BLV sera, stated as double positive or double negative by the combinationof commercial agar gel immunodiffusion (AGID) assay and a gp51-ELISA test, were tested with our in house dual rp24/rgp51 ELISA. Firstly, we checked the purified, optimized and standardized proteins as antigen by the checkerboard technique, and set up our in house ELISA test. The concordancecorrelation coefficient (CCC) and coefficient of variation (CV) intraplate repeatability levels were within the values established by the international standards.The statistical analysis demonstrated the value of sera correctly rankedhighest (93.48%), and for 0.3 cutoff, the sensitivity was 95.65% and the specificity 91.30%. In conclusion, the rp24/rgp51 ELISA developed and standardized here demonstrated to have good analytical characteristics to be considered for screening of BLV.
publishDate 2017
dc.date.none.fl_str_mv 2017-07
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/11336/49546
Larsen, Alejandra; Corva, Santiago; Panei, Carlos Javier; Geisler, Christoph; Mortola, Eduardo; Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease; Scientific Research Publishing Inc; Open Journal of Animal Sciences; 07; 03; 7-2017; 241-253
2161-7597
2161-7627
CONICET Digital
CONICET
url http://hdl.handle.net/11336/49546
identifier_str_mv Larsen, Alejandra; Corva, Santiago; Panei, Carlos Javier; Geisler, Christoph; Mortola, Eduardo; Enzyme-Linked Immunosorbent Assays Using the Recombinant gp51 and p24 of Bovine Leukemia Virus for Immunodetection of the Disease; Scientific Research Publishing Inc; Open Journal of Animal Sciences; 07; 03; 7-2017; 241-253
2161-7597
2161-7627
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.scirp.org/journal/doi.aspx?DOI=10.4236/ojas.2017.73019
info:eu-repo/semantics/altIdentifier/doi/10.4236/ojas.2017.73019
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
dc.publisher.none.fl_str_mv Scientific Research Publishing Inc
publisher.none.fl_str_mv Scientific Research Publishing Inc
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
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