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dc.contributor.author
Primo, Maria Evangelina  
dc.contributor.author
Thompson, Carolina Soledad  
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Valentini, Beatriz Susana  
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Sarli, Macarena  
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Novoa, María Belén  
dc.contributor.author
Mangold, Atilio Jose  
dc.contributor.author
de Echaide, Susana T.  
dc.date.available
2022-02-03T18:11:47Z  
dc.date.issued
2019-01  
dc.identifier.citation
Primo, Maria Evangelina; Thompson, Carolina Soledad; Valentini, Beatriz Susana; Sarli, Macarena; Novoa, María Belén; et al.; Development of a novel fusion protein with Anaplasma marginale and A. Centrale MSP5 improved performance of Anaplasma antibody detection by cELISA in infected and vaccinated cattle; Public Library of Science; Plos One; 14; 1; 1-2019; 1-13  
dc.identifier.issn
1932-6203  
dc.identifier.uri
http://hdl.handle.net/11336/151299  
dc.description.abstract
Detection of antibodies to Anaplasma spp. using commercial competitive enzyme-linked immunosorbent assay (ccELISA) is based on the recombinant major surface protein 5 fused to maltose binding protein (MBP-MSP5) or glutathione S-transferase (GST-MSP5). To avoid false positive reactions due to the presence of antibodies against E. coli MBP in cattle, previous sera absorption is required. This study evaluated the replacement of MBPMSP5 or GST-MSP5 antigens by the truncate MSP5 (residues 28–210) of A. marginale (tMSP5m), A. centrale (tMSP5c) and fusion protein MSP5 (tMSP5cm), expressed without N-terminus transmembrane helix in the ccELISA test. Immunoreactivity was evaluated by western blot using monoclonal antibodies against the tMSP5 and by in-house cELISA (hcELISA) with purified tMSP5m, tMSP5c or tMSP5cm using sera from cattle infected with A. marginale (n = 226) or vaccinated with A. centrale (n = 173) and uninfected cattle (n = 216). Results of hcELISA were compared with those of ccELISA. Recombinant protein was expressed highly soluble (> 95%) in E. coli without a molecular chaperone. Specificity of the hcELISA-tMSP5m, -MSP5c or -tMSP5cm was identical to (99.5%) and greater than that in ccELISA (96.3%). Sensitivity of hcELISA-tMSP5m and ccELISA was identical (95.5%), but lower than that of hcELISA-tMSP5cm (96.2%) and -tMSP5c (97.2%). The analysis of vaccinated cattle by hcELISA-tMSP5c showed sensitivity of 99.4%. In summary, the generation of fusion MSP5 A. marginale-A. centrale protein without transmembrane helix was a very effective method to express the recombinant protein highly soluble in the bacterial cytoplasm and contributed to an increased test performance for detecting antibodies in cattle naturally infected with A. marginale or vaccinated with A. centrale.  
dc.format
application/pdf  
dc.language.iso
eng  
dc.publisher
Public Library of Science  
dc.rights
info:eu-repo/semantics/openAccess  
dc.rights.uri
https://creativecommons.org/licenses/by/2.5/ar/  
dc.subject
ANAPLASMA MARGINALE  
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ANAPLASMA CENTRALE  
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ELISA  
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MSP5  
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Otras Ciencias Veterinarias  
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Ciencias Veterinarias  
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CIENCIAS AGRÍCOLAS  
dc.title
Development of a novel fusion protein with Anaplasma marginale and A. Centrale MSP5 improved performance of Anaplasma antibody detection by cELISA in infected and vaccinated cattle  
dc.type
info:eu-repo/semantics/article  
dc.type
info:ar-repo/semantics/artículo  
dc.type
info:eu-repo/semantics/publishedVersion  
dc.date.updated
2020-11-18T21:18:07Z  
dc.journal.volume
14  
dc.journal.number
1  
dc.journal.pagination
1-13  
dc.journal.pais
Estados Unidos  
dc.journal.ciudad
San Francisco  
dc.description.fil
Fil: Primo, Maria Evangelina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina  
dc.description.fil
Fil: Thompson, Carolina Soledad. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina  
dc.description.fil
Fil: Valentini, Beatriz Susana. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina  
dc.description.fil
Fil: Sarli, Macarena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina  
dc.description.fil
Fil: Novoa, María Belén. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina  
dc.description.fil
Fil: Mangold, Atilio Jose. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe; Argentina  
dc.description.fil
Fil: de Echaide, Susana T.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Santa Fe. Estación Experimental Agropecuaria Rafaela; Argentina  
dc.journal.title
Plos One  
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1371/journal.pone.0211149  
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0211149