Development and evaluation of a double-antigen sandwich ELISA to identify Anaplasma marginale–infected and A. centrale–vaccinated cattle
Bovine anaplasmosis is a worldwide infectious disease caused by the intraerythrocytic bacterium Anaplasma marginale, which is transmitted by ticks and fomites. A. centrale is a less virulent subspecies used as a live vaccine in cohorts of 8- to 10-mo-old calves that did not naturally reach enzooti...
| Main Authors: | , , , , , , , |
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| Format: | Artículo |
| Language: | Inglés |
| Published: |
American Association of Veterinary Laboratory Diagnosticians
2020
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| Subjects: | |
| Online Access: | http://hdl.handle.net/20.500.12123/7029 https://journals.sagepub.com/doi/10.1177/1040638719892953 https://doi.org/10.1177/1040638719892953 |
| _version_ | 1855483928537726976 |
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| author | Sarli, Macarena Thompson, Carolina Soledad Novoa, María Belen Valentini, Beatriz Susana Mastropaolo, Mariano Echaide, Ignacio Eduardo Torioni, Susana Marta Primo, María Evangelina |
| author_browse | Echaide, Ignacio Eduardo Mastropaolo, Mariano Novoa, María Belen Primo, María Evangelina Sarli, Macarena Thompson, Carolina Soledad Torioni, Susana Marta Valentini, Beatriz Susana |
| author_facet | Sarli, Macarena Thompson, Carolina Soledad Novoa, María Belen Valentini, Beatriz Susana Mastropaolo, Mariano Echaide, Ignacio Eduardo Torioni, Susana Marta Primo, María Evangelina |
| author_sort | Sarli, Macarena |
| collection | INTA Digital |
| description | Bovine anaplasmosis is a worldwide infectious disease caused by the intraerythrocytic bacterium Anaplasma
marginale, which is transmitted by ticks and fomites. A. centrale is a less virulent subspecies used as a live vaccine in
cohorts of 8- to 10-mo-old calves that did not naturally reach enzootic stability. We developed 3 variants of a double-antigen
sandwich ELISA (dasELISA) using a recombinant major surface protein 5 (MSP5) from A. marginale (dasELISAm) or
from A. centrale (dasELISAc) or using MSP5 from both organisms (dasELISAmc). Each dasELISA was tested for the
detection of antibodies against A. marginale and A. centrale. The tests were validated using serum samples from cattle not
infected with Anaplasma spp. (n = 388), infected with A. marginale (n = 436), and vaccinated with A. centrale (n = 358),
confirmed by nested PCR. A total of 462 samples were compared with a commercial competitive ELISA (cELISA). For
dasELISAm, dasELISAc, and dasELISAmc, specificities were 98.7%, 98.7%, and 97.4%, and overall sensitivities were
92.6%, 85.7%, and 97.4%, respectively. For A. marginale–infected and A. centrale–vaccinated cattle, sensitivities were
97.7% and 86.3% for dasELISAm, and 77.7% and 95.5% for dasELISAc, respectively. Sensitivity of dasELISAmc was
similar for both groups (>96%). The agreement rate between dasELISAmc and cELISA was 96.3% (κ = 0.92); the former
test allowed earlier detection of seroconversion of vaccinated cattle than did cELISA. Based on these results, the test could
be used to 1) determine the enzootic stability or instability of anaplasmosis in calves, 2) conduct epidemiologic studies, and
3) evaluate the immunogenicity of A. centrale live vaccine. |
| format | Artículo |
| id | INTA7029 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2020 |
| publishDateRange | 2020 |
| publishDateSort | 2020 |
| publisher | American Association of Veterinary Laboratory Diagnosticians |
| publisherStr | American Association of Veterinary Laboratory Diagnosticians |
| record_format | dspace |
| spelling | INTA70292023-04-26T17:04:02Z Development and evaluation of a double-antigen sandwich ELISA to identify Anaplasma marginale–infected and A. centrale–vaccinated cattle Sarli, Macarena Thompson, Carolina Soledad Novoa, María Belen Valentini, Beatriz Susana Mastropaolo, Mariano Echaide, Ignacio Eduardo Torioni, Susana Marta Primo, María Evangelina Anaplasma Marginale ELISA Anaplasmosis Enfermedades Infecciosas Bovinae Infectious Diseases Anaplasma centrale Double-antigen Sandwich ELISA Bovine anaplasmosis is a worldwide infectious disease caused by the intraerythrocytic bacterium Anaplasma marginale, which is transmitted by ticks and fomites. A. centrale is a less virulent subspecies used as a live vaccine in cohorts of 8- to 10-mo-old calves that did not naturally reach enzootic stability. We developed 3 variants of a double-antigen sandwich ELISA (dasELISA) using a recombinant major surface protein 5 (MSP5) from A. marginale (dasELISAm) or from A. centrale (dasELISAc) or using MSP5 from both organisms (dasELISAmc). Each dasELISA was tested for the detection of antibodies against A. marginale and A. centrale. The tests were validated using serum samples from cattle not infected with Anaplasma spp. (n = 388), infected with A. marginale (n = 436), and vaccinated with A. centrale (n = 358), confirmed by nested PCR. A total of 462 samples were compared with a commercial competitive ELISA (cELISA). For dasELISAm, dasELISAc, and dasELISAmc, specificities were 98.7%, 98.7%, and 97.4%, and overall sensitivities were 92.6%, 85.7%, and 97.4%, respectively. For A. marginale–infected and A. centrale–vaccinated cattle, sensitivities were 97.7% and 86.3% for dasELISAm, and 77.7% and 95.5% for dasELISAc, respectively. Sensitivity of dasELISAmc was similar for both groups (>96%). The agreement rate between dasELISAmc and cELISA was 96.3% (κ = 0.92); the former test allowed earlier detection of seroconversion of vaccinated cattle than did cELISA. Based on these results, the test could be used to 1) determine the enzootic stability or instability of anaplasmosis in calves, 2) conduct epidemiologic studies, and 3) evaluate the immunogenicity of A. centrale live vaccine. EEA Rafaela Fil: Sarli, Macarena. 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: Novoa, María Belen. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fil: Valentini, Beatriz Susana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina Fil: Mastropaolo, Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias; Argentina Fil: Echaide, Ignacio Eduardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina. Fil: Torioni de Echaide, Susana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina. Fil: Primo, María Evangelina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. 2020-04-02T14:54:43Z 2020-04-02T14:54:43Z 2019-11 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/7029 https://journals.sagepub.com/doi/10.1177/1040638719892953 1040-6387 1943-4936 (Online) https://doi.org/10.1177/1040638719892953 eng info:eu-repo/semantics/openAccess application/pdf American Association of Veterinary Laboratory Diagnosticians Journal of veterinary diagnostic investigation 32 (1) : 70-76. (January 1, 2020) |
| spellingShingle | Anaplasma Marginale ELISA Anaplasmosis Enfermedades Infecciosas Bovinae Infectious Diseases Anaplasma centrale Double-antigen Sandwich ELISA Sarli, Macarena Thompson, Carolina Soledad Novoa, María Belen Valentini, Beatriz Susana Mastropaolo, Mariano Echaide, Ignacio Eduardo Torioni, Susana Marta Primo, María Evangelina Development and evaluation of a double-antigen sandwich ELISA to identify Anaplasma marginale–infected and A. centrale–vaccinated cattle |
| title | Development and evaluation of a double-antigen sandwich ELISA to identify Anaplasma marginale–infected and A. centrale–vaccinated cattle |
| title_full | Development and evaluation of a double-antigen sandwich ELISA to identify Anaplasma marginale–infected and A. centrale–vaccinated cattle |
| title_fullStr | Development and evaluation of a double-antigen sandwich ELISA to identify Anaplasma marginale–infected and A. centrale–vaccinated cattle |
| title_full_unstemmed | Development and evaluation of a double-antigen sandwich ELISA to identify Anaplasma marginale–infected and A. centrale–vaccinated cattle |
| title_short | Development and evaluation of a double-antigen sandwich ELISA to identify Anaplasma marginale–infected and A. centrale–vaccinated cattle |
| title_sort | development and evaluation of a double antigen sandwich elisa to identify anaplasma marginale infected and a centrale vaccinated cattle |
| topic | Anaplasma Marginale ELISA Anaplasmosis Enfermedades Infecciosas Bovinae Infectious Diseases Anaplasma centrale Double-antigen Sandwich ELISA |
| url | http://hdl.handle.net/20.500.12123/7029 https://journals.sagepub.com/doi/10.1177/1040638719892953 https://doi.org/10.1177/1040638719892953 |
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