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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/49546
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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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1846083261226287104 |
score |
13.22299 |