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Detection of anti-Neospora caninum antibodies in sheep’s full-cream milk by time-resolved fluorescence immunoassay

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dc.contributor.authorHuertas-López, Ana
dc.contributor.authorSánchez-Sánchez, Roberto
dc.contributor.authorDiezma-Díaz, Carlos
dc.contributor.authorAlvarez-García, Gema
dc.contributor.authorMartínez-Carrasco Pleite, Carlos
dc.contributor.authorMartínez Subiela, Silvia
dc.contributor.authorCerón Madrigal, José Joaquín
dc.contributor.departmentSanidad Animal
dc.contributor.otherFacultad de Veterinaria
dc.date.accessioned2025-12-28T10:18:46Z
dc.date.available2025-12-28T10:18:46Z
dc.date.copyright© 2021 The Authors
dc.date.issued2021-12-25
dc.description.abstractOvine neosporosis, caused by the Apicomplexan parasite Neospora caninum, leads to reproductive failure worldwide. Nowadays, there is a trend to develop diagnostic techniques using non-invasive samples, such as milk, in order to reduce animal stress, sample collection effort, and costs. The objective of this study was to develop and validate a highly sensitive and specific serological technique, based on a time resolved-fluorescence immunoassay using a N. caninum GRA7 antigen (GRA7-TRFIA), for the detection of anti-N. caninum immunoglobulins G on sheep’ full-cream milk samples. An analytical validation was performed, including intra- and inter-assay precision, analytical sensitivity and accuracy. The diagnostic performance of the assay was evaluated by studying the positive-negative discrimination by Mann Whitney U tests. In additon optimal cut-offs, diagnostic sensitivity and specificity, and areas under the curve were calculated by three Receiver Operating Curve (ROC) analyses, using GRA7-TRFIA and a N. caninum tachyzoite soluble extract-based ELISA (NcSALUVET-ELISA) in blood sera, and the coinciding results of both techniques, as reference techniques. Moreover, Spearman’s correlation of GRA7-TRFIA in milk with the techniques in sera and agreement (kappa values) were also estimated. GRA7-TRFIA for milk samples showed an adequate precision, with high analytical sensitivity and accuracy. Regarding ROC analyses, at the optimal cut-offs, the diagnostic sensitivity and specificity were more than 90 % in all cases. In addition, GRA7-TRFIA values in milk were more positively correlated to GRA7-TRFIA values in blood sera than in the case of values obtained with NcSALUVET-ELISA. GRA7-TRFIA in milk showed an almost perfect agreement with GRA7-TRFIA in blood sera (kappa = 0.98) and with the coinciding results of GRA7-TRFIA and NcSALUVET in blood sera (kappa = 1.00), while it has a substantial agreement with NcSALUVET-ELISA (kappa = 0.69). In the light of these results, GRA7-TRFIA in full-cream milk samples is a highly sensitive technique that could be used for screening anti-N. caninum antibodies in sheep flocks.
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dc.format.extent9
dc.identifier.citationVeterinary Parasitology 301 (2022) 109641
dc.identifier.doihttps://doi.org/10.1016/j.vetpar.2021.109641
dc.identifier.eissn1873-2550
dc.identifier.issn0304-4017
dc.identifier.urihttp://hdl.handle.net/10201/182889
dc.languageeng
dc.publisherElsevier
dc.relationAna Huertas López was supported by a pre-doctoral grant from the University of Murcia (R-1207/2017). Roberto Sánchez Sánchez was supported by the Community of Madrid, Spain (PLATESA2, S2018/BAA-430). Gema ´Alvarez García is part of the TOXOSOURCES consortium, supported by funding from the European Union’s Horizon 2020 Research and Innovation Programme under grant agreement No. 773830: One Health European Joint Programme. This work was supported by the Seneca Foundation of Murcia Region, Spain (project 19,894/GERM/15).
dc.relation.publisherversionhttps://www.sciencedirect.com/science/article/pii/S0304401721003010?via%3Dihub
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 International*
dc.rights.accessRightsinfo:eu-repo/semantics/openAccess
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subjectFull-cream milk
dc.subjectSheep
dc.subjectTime-resolved fluorescence immunoassay
dc.subjectSerology
dc.subjectNeospora caninum
dc.subject.odsObjetivo 3: Salud
dc.titleDetection of anti-Neospora caninum antibodies in sheep’s full-cream milk by time-resolved fluorescence immunoassay
dc.typeinfo:eu-repo/semantics/article
dc.type.versioninfo:eu-repo/semantics/publishedVersión
dspace.entity.typePublicationes
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relation.isAuthorOfPublication.latestForDiscovery74bbaf1b-2a73-4ec8-b1c6-429311688d3b
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