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
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 pre...
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| Format: | Artículo |
| Language: | Inglés |
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Plos One
2019
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| Subjects: | |
| Online Access: | https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0211149 http://hdl.handle.net/20.500.12123/4751 https://doi.org/10.1371/journal.pone.0211149 |
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| author | Primo, María Evangelina Thompson, Carolina Soledad Valentini, Beatriz Susana Sarli, Macarena Novoa, María Belen Mangold, Atilio Jose Torioni, Susana Marta |
| author_browse | Mangold, Atilio Jose Novoa, María Belen Primo, María Evangelina Sarli, Macarena Thompson, Carolina Soledad Torioni, Susana Marta Valentini, Beatriz Susana |
| author_facet | Primo, María Evangelina Thompson, Carolina Soledad Valentini, Beatriz Susana Sarli, Macarena Novoa, María Belen Mangold, Atilio Jose Torioni, Susana Marta |
| author_sort | Primo, María Evangelina |
| collection | INTA Digital |
| description | 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 MBP-MSP5 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. |
| format | Artículo |
| id | INTA4751 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2019 |
| publishDateRange | 2019 |
| publishDateSort | 2019 |
| publisher | Plos One |
| publisherStr | Plos One |
| record_format | dspace |
| spelling | INTA47512019-03-27T12:05:00Z 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 Primo, María Evangelina Thompson, Carolina Soledad Valentini, Beatriz Susana Sarli, Macarena Novoa, María Belen Mangold, Atilio Jose Torioni, Susana Marta Anaplasma Marginale Anticuerpos Ganado Bovino ELISA Enfermedades de los Animales Anaplasmosis Proteínas Antibodies Cattle Animal Diseases Proteins Anaplasma Centrale 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 MBP-MSP5 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. EEA Rafaela Fil: Primo, María Evangelina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Thompson, Carolina Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina Fil: Valentini, Beatriz Susana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina Fil: Sarli, Macarena. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Novoa, María Belen. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Mangold, Atilio Jose. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Torioni, Susana Marta. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina 2019-03-27T11:31:45Z 2019-03-27T11:31:45Z 2019-01-23 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0211149 http://hdl.handle.net/20.500.12123/4751 1932-6203 https://doi.org/10.1371/journal.pone.0211149 eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Plos One PLoS ONE 14 (1): e0211149. (January 2019) |
| spellingShingle | Anaplasma Marginale Anticuerpos Ganado Bovino ELISA Enfermedades de los Animales Anaplasmosis Proteínas Antibodies Cattle Animal Diseases Proteins Anaplasma Centrale Primo, María Evangelina Thompson, Carolina Soledad Valentini, Beatriz Susana Sarli, Macarena Novoa, María Belen Mangold, Atilio Jose Torioni, Susana Marta 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 |
| 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 |
| title_full | 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 |
| title_fullStr | 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 |
| title_full_unstemmed | 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 |
| title_short | 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 |
| title_sort | 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 |
| topic | Anaplasma Marginale Anticuerpos Ganado Bovino ELISA Enfermedades de los Animales Anaplasmosis Proteínas Antibodies Cattle Animal Diseases Proteins Anaplasma Centrale |
| url | https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0211149 http://hdl.handle.net/20.500.12123/4751 https://doi.org/10.1371/journal.pone.0211149 |
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