RT Journal Article
T1 Detection of anti-Neospora caninum antibodies in sheep’s full-cream milk by a time-resolved fluorescence immunoassay
A1 Huertas Lopez, Ana
A1 Sánchez Sánchez, Roberto
A1 Diezma Díaz, Carlos
A1 Álvarez García, Gema
A1 Martínez Carrasco, Carlos
A1 Martínez Subiela, Silvia
A1 Cerón, José Joaquín
AB Ovine 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.
PB Elsevier
SN 0304-4017
YR 2022
FD 2022
LK https://hdl.handle.net/20.500.14352/111995
UL https://hdl.handle.net/20.500.14352/111995
LA eng
NO Huertas-López, A., Sánchez-Sánchez, R., Diezma-Díaz, C., Álvarez-García, G., Martínez-Carrasco, C., Martínez-Subiela, S., & Cerón, J. J. (2022). Detection of anti-Neospora caninum antibodies in sheep's full-cream milk by a time-resolved fluorescence immunoassay. Veterinary parasitology, 301, 109641. https://doi.org/10.1016/j.vetpar.2021.109641
NO JUSTIFICACIÓN DE AUTORES:Huertas-López, Ana: Conceptualization; Formal analysis; Investigation; Methodology; Validation; Roles/Writing - original draft; Writing - review & editing.Sánchez-Sánchez, Roberto: Data curation; Investigation; Methodology; Writing - review & editing.Diezma-Díaz, Carlos: Data curation; Investigation; Methodology; Writing - review & editing.Álvarez-García, Gema: Conceptualization; Funding acquisition; Project administration; Supervision; Validation; Visualization; Writing - review & editing.Martínez-Carrasco, Carlos: Conceptualization; Project administration; Supervision; Validation; Visualization; Writing - review & editing.Martínez-Subiela, Silvia: Conceptualization; Formal analysis; Investigation; Methodology; Software; Supervision; Validation; Visualization; Writing - review & editing.Cerón, José Joaquín: Conceptualization; Funding acquisition; Project administration; Supervision; Validation; Visualization; Writing - review & editing.
NO Universidad de Murcia
NO Comunidad de Madrid
NO European Commission
NO Fundación Séneca
DS Docta Complutense
RD 7 abr 2025