Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale
A species-specific competitive ELISA for Anaplasma marginale antibodies detection (cELISA-sAm) was developed by using both a monoclonal antibody specific to an epitope of MSP5 of A. marginale (Am6-mAb) and adding a pre-incubation phase with the truncated MSP5 of A. centrale (tMSP5c). For the determi...
| Main Authors: | , , , , |
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| Format: | Artículo |
| Language: | Inglés |
| Published: |
Elsevier
2026
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| Online Access: | http://hdl.handle.net/20.500.12123/24969 https://www.sciencedirect.com/science/article/abs/pii/S1090023325002321 https://doi.org/10.1016/j.tvjl.2025.106528 |
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| author | Bellezze, Julio Valentini, Beatriz Susana Bosio, Anabela Thompson, Carolina Soledad Primo, María Evangelina |
| author_browse | Bellezze, Julio Bosio, Anabela Primo, María Evangelina Thompson, Carolina Soledad Valentini, Beatriz Susana |
| author_facet | Bellezze, Julio Valentini, Beatriz Susana Bosio, Anabela Thompson, Carolina Soledad Primo, María Evangelina |
| author_sort | Bellezze, Julio |
| collection | INTA Digital |
| description | A species-specific competitive ELISA for Anaplasma marginale antibodies detection (cELISA-sAm) was developed by using both a monoclonal antibody specific to an epitope of MSP5 of A. marginale (Am6-mAb) and adding a pre-incubation phase with the truncated MSP5 of A. centrale (tMSP5c). For the determination of the cutoff value, diagnostic sensitivity (DSe) and specificity (DSp) of cELISA-sAm, 382 samples from cattle not infected with Anaplasma spp., 237 samples from cattle naturally infected with A. marginale, and 102 samples from cattle vaccinated with A. centrale were analyzed. The infection status was established using nested PCR (nPCR) or nPCR-restriction fragment length polymorphism assay (nPCR-RFLP) as reference technique. In a field evaluation, cELISA-sAm and nPCR-RFLP were used on Anaplasma spp. positive samples by double-antigen sandwich ELISA (dasELISA) from 2 herds to confirm the A. marginale infection. The cELISA-sAm optimal cutoff value was ≥ 18 %I, with a DSe of 89.9 % and a DSp of 99.0 %. In a field evaluation, concordance between cELISA-sAm and nPCR-RFLP was 93.0 % with κ = 0.81 (95 % CI = 0.56–1.00) for A. marginale-infected cattle detection. In countries in which prevention is based on vaccination with A. centrale, the replacement of current ELISAs by this cELISA-sAm would decrease the false positive results and avoid the need of species confirmation by molecular techniques when disease control programs are performed to monitor the prevalence of A. marginale. |
| format | Artículo |
| id | INTA24969 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2026 |
| publishDateRange | 2026 |
| publishDateSort | 2026 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA249692026-01-09T14:50:10Z Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale Bellezze, Julio Valentini, Beatriz Susana Bosio, Anabela Thompson, Carolina Soledad Primo, María Evangelina Anaplasma marginale ELISA Antibodies Cattle Immune Response Monoclonal Antibodies Anticuerpos Ganado Bovino Respuesta Inmunológica Anticuerpos Monoclonales A species-specific competitive ELISA for Anaplasma marginale antibodies detection (cELISA-sAm) was developed by using both a monoclonal antibody specific to an epitope of MSP5 of A. marginale (Am6-mAb) and adding a pre-incubation phase with the truncated MSP5 of A. centrale (tMSP5c). For the determination of the cutoff value, diagnostic sensitivity (DSe) and specificity (DSp) of cELISA-sAm, 382 samples from cattle not infected with Anaplasma spp., 237 samples from cattle naturally infected with A. marginale, and 102 samples from cattle vaccinated with A. centrale were analyzed. The infection status was established using nested PCR (nPCR) or nPCR-restriction fragment length polymorphism assay (nPCR-RFLP) as reference technique. In a field evaluation, cELISA-sAm and nPCR-RFLP were used on Anaplasma spp. positive samples by double-antigen sandwich ELISA (dasELISA) from 2 herds to confirm the A. marginale infection. The cELISA-sAm optimal cutoff value was ≥ 18 %I, with a DSe of 89.9 % and a DSp of 99.0 %. In a field evaluation, concordance between cELISA-sAm and nPCR-RFLP was 93.0 % with κ = 0.81 (95 % CI = 0.56–1.00) for A. marginale-infected cattle detection. In countries in which prevention is based on vaccination with A. centrale, the replacement of current ELISAs by this cELISA-sAm would decrease the false positive results and avoid the need of species confirmation by molecular techniques when disease control programs are performed to monitor the prevalence of A. marginale. EEA Rafaela Fil: Bellezze, Julio. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Bellezze, Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Valentini, Beatriz Susana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Valentini, Beatriz Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Bosio, Anabela. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Bosio, Anabela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Thompson, Carolina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Thompson, Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Primo, María Evangelina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Primo, María Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina 2026-01-09T14:46:19Z 2026-01-09T14:46:19Z 2026-02 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/24969 https://www.sciencedirect.com/science/article/abs/pii/S1090023325002321 1090-0233 1532-2971 https://doi.org/10.1016/j.tvjl.2025.106528 eng info:eu-repo/semantics/restrictedAccess 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 Elsevier The Veterinary Journal 315 : 106528. (February 2026) |
| spellingShingle | Anaplasma marginale ELISA Antibodies Cattle Immune Response Monoclonal Antibodies Anticuerpos Ganado Bovino Respuesta Inmunológica Anticuerpos Monoclonales Bellezze, Julio Valentini, Beatriz Susana Bosio, Anabela Thompson, Carolina Soledad Primo, María Evangelina Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale |
| title | Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale |
| title_full | Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale |
| title_fullStr | Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale |
| title_full_unstemmed | Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale |
| title_short | Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale |
| title_sort | development and field evaluation of a competitive elisa to detect specific antibodies against anaplasma marginale |
| topic | Anaplasma marginale ELISA Antibodies Cattle Immune Response Monoclonal Antibodies Anticuerpos Ganado Bovino Respuesta Inmunológica Anticuerpos Monoclonales |
| url | http://hdl.handle.net/20.500.12123/24969 https://www.sciencedirect.com/science/article/abs/pii/S1090023325002321 https://doi.org/10.1016/j.tvjl.2025.106528 |
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