{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,10]],"date-time":"2026-03-10T04:35:20Z","timestamp":1773117320960,"version":"3.50.1"},"reference-count":20,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2018,1,23]],"date-time":"2018-01-23T00:00:00Z","timestamp":1516665600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"PICT","award":["454 - 2011"],"award-info":[{"award-number":["454 - 2011"]}]},{"name":"PICT","award":["932 - 2011"],"award-info":[{"award-number":["932 - 2011"]}]},{"name":"PNBIO","award":["1131032"],"award-info":[{"award-number":["1131032"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Veterinary Sciences"],"abstract":"<jats:p>The current method for Babesia spp. serodiagnosis based on a crude merozoite antigen is a complex and time-consuming procedure. An indirect enzyme-linked immunosorbent assay (iELISA) based on a recombinant multi-antigen of Babesia bovis (rMABbO) was developed for detection of antibodies in bovines suspected of infection with this parasite. The multi-antigen comprises gene fragments of three previously characterized B. bovis antigens: MSA-2c, RAP-1 and the Heat Shock protein 20 that are well-conserved among geographically distant strains. The cutoff value for the new rMABbo-iELISA was determined using 75 known\u2014positive and 300 known\u2014negative bovine sera previously tested for antibodies to B. bovis by the gold-standard ELISA which uses a merozoite lysate. A cutoff value of \u226535% was determined in these samples by receiver operator characteristic (ROC) curve analysis, showing a sensitivity of 95.9% and a specificity of 94.3%. The rMABbo-iELISA was further tested in a blind trial using an additional set of 263 field bovine sera from enzootic and tick-free regions of Argentina. Results showed a good agreement with the gold standard test with a Cohen\u2019s kappa value of 0.76. Finally, the prevalence of bovine babesiosis in different tick enzootic regions of Argentina was analyzed where seropositivity values among 68\u201380% were obtained. A certain level of cross reaction was observed when samples from B. bigemina infected cattle were analyzed with the new test, which can be attributed to shared epitopes between 2 of the 3 antigens. This new rMABbo-iELISA could be considered a simpler alternative to detect anti Babesia spp. antibodies and appears to be well suited to perform epidemiological surveys at the herd level in regions where ticks are present.<\/jats:p>","DOI":"10.3390\/vetsci5010013","type":"journal-article","created":{"date-parts":[[2018,1,23]],"date-time":"2018-01-23T13:06:51Z","timestamp":1516712811000},"page":"13","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Development of an Indirect ELISA Based on a Recombinant Chimeric Protein for the Detection of Antibodies against Bovine Babesiosis"],"prefix":"10.3390","volume":"5","author":[{"given":"Jos\u00e9","family":"Jaramillo Ortiz","sequence":"first","affiliation":[{"name":"Instituto de Biotecnolog\u00eda, Centro de Investigaciones en Ciencias Veterinarias y Agron\u00f3micas, Instituto Nacional de Tecnolog\u00eda Agropecuaria, De Los Reseros y Dr. Nicol\u00e1s Repetto s\/no., Hurlingham 1686, Buenos Aires, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Valeria","family":"Montenegro","sequence":"additional","affiliation":[{"name":"Instituto de Biotecnolog\u00eda, Centro de Investigaciones en Ciencias Veterinarias y Agron\u00f3micas, Instituto Nacional de Tecnolog\u00eda Agropecuaria, De Los Reseros y Dr. Nicol\u00e1s Repetto s\/no., Hurlingham 1686, Buenos Aires, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sof\u00eda","family":"De la Fourni\u00e8re","sequence":"additional","affiliation":[{"name":"Instituto de Biotecnolog\u00eda, Centro de Investigaciones en Ciencias Veterinarias y Agron\u00f3micas, Instituto Nacional de Tecnolog\u00eda Agropecuaria, De Los Reseros y Dr. Nicol\u00e1s Repetto s\/no., Hurlingham 1686, Buenos Aires, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"N\u00e9stor","family":"Sarmiento","sequence":"additional","affiliation":[{"name":"Estaci\u00f3n Experimental Agropecuaria Mercedes, Instituto Nacional de Tecnolog\u00eda Agropecuaria, Juan Pujol al Este, s\/n, Corrientes CP 3470, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marisa","family":"Farber","sequence":"additional","affiliation":[{"name":"Instituto de Biotecnolog\u00eda, Centro de Investigaciones en Ciencias Veterinarias y Agron\u00f3micas, Instituto Nacional de Tecnolog\u00eda Agropecuaria, De Los Reseros y Dr. Nicol\u00e1s Repetto s\/no., Hurlingham 1686, Buenos Aires, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Silvina","family":"Wilkowsky","sequence":"additional","affiliation":[{"name":"Instituto de Biotecnolog\u00eda, Centro de Investigaciones en Ciencias Veterinarias y Agron\u00f3micas, Instituto Nacional de Tecnolog\u00eda Agropecuaria, De Los Reseros y Dr. Nicol\u00e1s Repetto s\/no., Hurlingham 1686, Buenos Aires, Argentina"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2018,1,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"S247","DOI":"10.1017\/S0031182004005190","article-title":"Babesiosis of cattle","volume":"129","author":"Bock","year":"2004","journal-title":"Parasitology"},{"key":"ref_2","first-page":"116","article-title":"Estimating of direct economic losses caused by the tick Boophilus microplus and associated diseases in Argentina","volume":"23","author":"Guglielmone","year":"1994","journal-title":"Therios"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/S0169-4758(99)01471-4","article-title":"Designing blood-stage vaccines against Babesia bovis and B. bigemina","volume":"15","author":"Brown","year":"1999","journal-title":"Parasitol. Today"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Florin-christensen, M., Suarez, C.E., Rodriguez, A.E., Flores, D.A., and Schnittger, L. (2014). Vaccines against bovine babesiosis: Where we are now and possible roads ahead. Parasitol. Today, 1\u201330.","DOI":"10.1017\/S0031182014000961"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1016\/j.vetpar.2003.07.011","article-title":"Attainment of endemic stability to Babesia bigemina in cattle on a South African ranch where non-intensive tick control was applied","volume":"116","author":"Regassa","year":"2003","journal-title":"Vet. Parasitol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1196\/annals.1307.025","article-title":"Use of a monoclonal antibody against Babesia bovis Merozoite Surface Antigen-2c for the development of a competitive ELISA test","volume":"1026","author":"Dominguez","year":"2004","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"924","DOI":"10.1128\/CVI.00015-12","article-title":"Validation and field evaluation of a competitive enzyme-linked immunosorbent assay for diagnosis of Babesia bovis infections in Argentina","volume":"19","author":"Dominguez","year":"2012","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3371","DOI":"10.1128\/JCM.40.10.3771-3775.2002","article-title":"Evaluation of an enzyme\u2014Linked immunosorbent assay with recombinant rhoptry-associated protein 1 antigen against Babesia bovis for the detection of specific antibodies in cattle","volume":"40","author":"Boonchit","year":"2002","journal-title":"J. Clin. Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1016\/j.vetpar.2008.07.025","article-title":"Efficiency of a recombinant MSA-2c-based ELISA to establish the persistence of antibodies in cattle vaccinated with Babesia bovis","volume":"157","author":"Bono","year":"2008","journal-title":"Vet. Parasitol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3913","DOI":"10.1016\/j.vaccine.2016.05.053","article-title":"Evaluation of different heterologous prime-boost immunization strategies against Babesia bovis using viral vectored and protein-adjuvant vaccines based on a chimeric multi-antigen","volume":"34","author":"Molinari","year":"2016","journal-title":"Vaccine"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1016\/0167-5877(95)00485-F","article-title":"Evaluation of an enzyme-linked immunosorbent assay kit to detect Babesia bovis in cattle","volume":"24","author":"Echaide","year":"1995","journal-title":"Prev. Vet. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"6576","DOI":"10.1016\/j.vaccine.2014.06.056","article-title":"Influence of antibodies transferred by colostrum in the immune responses of calves to current foot-and-mouth disease vaccines","volume":"12","author":"Bucafusco","year":"2014","journal-title":"Vaccine"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.vetpar.2014.01.009","article-title":"Veterinary Parasitology Diagnostic value of the recombinant tandem repeat antigen TeGM6-4r for surra in water buffaloes","volume":"201","author":"Nguyen","year":"2014","journal-title":"Vet. Parasitol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Altman, D. (1991). Practical Statistics for Medical Research, Chapman Hall.","DOI":"10.1201\/9780429258589"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1504","DOI":"10.2174\/092986712799828355","article-title":"Current Advances in Detection and Treatment of Babesiosis","volume":"19","author":"Mosqueda","year":"2012","journal-title":"Curr. Med. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"139","DOI":"10.1016\/j.vetpar.2014.05.010","article-title":"Validation of a recombinant protein indirect ELISA for the detection of specific antibodies against Theileria uilenbergi and Theileria luwenshuni in small ruminants","volume":"204","author":"Liu","year":"2014","journal-title":"Vet. Parasitol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1007\/s00436-016-5332-7","article-title":"Evaluating an indirect rMPSP enzyme-linked immunosorbent assay for the detection of bovine Theileria infection in China","volume":"116","author":"Zhao","year":"2016","journal-title":"Parasitol. Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/0166-6851(91)90197-E","article-title":"Characterization of the gene encoding a 60-kilodalton Babesia bovis merozoite protein with conserved and surface-exposed epitopes","volume":"46","author":"Suarez","year":"1991","journal-title":"Mol. Biochem. Parasitol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/S0166-6851(01)00375-9","article-title":"A novel 20-kilodalton protein conserved in Babesia bovis and B. bigemina stimulates memory CD4(+) T lymphocyte responses in B. bovis-immune cattle","volume":"1","author":"Brown","year":"2001","journal-title":"Mol. Biochem. Parasitol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Pillay, D.I., Zotte, J., Fikru, R., B\u00fcscher, P., Mucache, H., and Neves, L. (2013). Trypanosoma vivax GM6 Antigen: A Candidate Antigen for Diagnosis of African Animal Trypanosomosis in Cattle. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0078565"}],"container-title":["Veterinary Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2306-7381\/5\/1\/13\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T14:52:19Z","timestamp":1760194339000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2306-7381\/5\/1\/13"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,1,23]]},"references-count":20,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2018,3]]}},"alternative-id":["vetsci5010013"],"URL":"https:\/\/doi.org\/10.3390\/vetsci5010013","relation":{},"ISSN":["2306-7381"],"issn-type":[{"value":"2306-7381","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,1,23]]}}}