Evaluation of species-specific polyclonal antibodies to detect and differentiate between Neospora caninum and Toxoplasma gondii

Autores
Lepore, Tanja; Macrae, Alastair I.; Canton, German Jose; Cantile, Carlo; Martineau, Henny M.; Palarea-Albaladejo, Javier; Cahalan, Stephen; Underwood, Clare; Katzer, Frank; Chianini, Francesca
Año de publicación
2024
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Neosporosis and toxoplasmosis are major causes of abortion in livestock worldwide, leading to substantial economic losses. Detection tools are fundamental to the diagnosis and management of those diseases. Current immunohistochemistry (IHC) tests, using sera raised against whole parasite lysates, have not been able to distinguish between Toxoplasma gondii and Neospora caninum. We used T. gondii and N. caninum recombinant proteins, expressed in Escherichia coli and purified using insoluble conditions, to produce specific polyclonal rabbit antisera. We aimed to develop species-specific sera that could be used in IHC on formalin-fixed, paraffin-embedded (FFPE) tissue sections to improve the diagnosis of ruminant abortions caused by protozoa. Two polyclonal rabbit sera, raised against recombinant proteins, anti–Neospora-rNcSRS2 and anti–Toxoplasma-rTgSRS2, had specificity for the parasite they were raised against. We tested the specificity for each polyclonal serum using FFPE tissue sections known to be infected with T. gondii and N. caninum. The anti–Neospora-rNcSRS2 serum labeled specifically only N. caninum–infected tissue blocks, and the anti–Toxoplasma-rTgSRS2 serum was specific to only T. gondii–infected tissues. Moreover, tissues from 52 cattle and 19 sheep previously diagnosed by lesion profiles were tested using IHC with our polyclonal sera and PCR. The overall agreement between IHC and PCR was 90.1% for both polyclonal anti-rNcSRS2 and anti-rTgSRS2 sera. The polyclonal antisera were specific and allowed visual confirmation of protozoan parasites by IHC, but they were not as sensitive as PCR testing.
EEA Balcarce
Fil: Lepore, Tanja. Moredun Research Institute; Reino Unido
Fil: Macrae, Alastair I. Royal School of Veterinary Studies; Reino Unido
Fil: Cantón, Germán. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina
Fil: Cantile, Carlo. Department of Veterinary Sciences; Italia
Fil: Martineau, Henny M. Royal Veterinary College. Department of Infectious Diseases; Reino Unido
Fil: Palarea-Albaladejo, Javier. Biomathematics and Statistics Scotland; Reino Unido
Fil: Cahalan, Stephen. Royal Veterinary College. Department of Infectious Diseases; Reino Unido
Fil: Underwood, Clare. Moredun Research Institute; Reino Unido
Fil: Katzer, Frank. Moredun Research Institute; Reino Unido
Fil: Chianin, Francesca. Moredun Research Institute; Reino Unido
Fuente
Journal of Veterinary Diagnostic Investigation 36 (3) : 418-427 (2024)
Materia
Neospora caninum
Toxoplasma gondii
Neosporosi
Toxoplasmosis
Aborto
Ganado
Anticuerpos Policlonales
Neosporosis
Abortion
Livestock
Polyclonal Antibodies
Nivel de accesibilidad
acceso restringido
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/27463
Acceder
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oai_identifier_str oai:localhost:20.500.12123/27463
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network_name_str INTA Digital (INTA)
spelling Evaluation of species-specific polyclonal antibodies to detect and differentiate between Neospora caninum and Toxoplasma gondiiLepore, TanjaMacrae, Alastair I.Canton, German JoseCantile, CarloMartineau, Henny M.Palarea-Albaladejo, JavierCahalan, StephenUnderwood, ClareKatzer, FrankChianini, FrancescaNeospora caninumToxoplasma gondiiNeosporosiToxoplasmosisAbortoGanadoAnticuerpos PoliclonalesNeosporosisAbortionLivestockPolyclonal AntibodiesNeosporosis and toxoplasmosis are major causes of abortion in livestock worldwide, leading to substantial economic losses. Detection tools are fundamental to the diagnosis and management of those diseases. Current immunohistochemistry (IHC) tests, using sera raised against whole parasite lysates, have not been able to distinguish between Toxoplasma gondii and Neospora caninum. We used T. gondii and N. caninum recombinant proteins, expressed in Escherichia coli and purified using insoluble conditions, to produce specific polyclonal rabbit antisera. We aimed to develop species-specific sera that could be used in IHC on formalin-fixed, paraffin-embedded (FFPE) tissue sections to improve the diagnosis of ruminant abortions caused by protozoa. Two polyclonal rabbit sera, raised against recombinant proteins, anti–Neospora-rNcSRS2 and anti–Toxoplasma-rTgSRS2, had specificity for the parasite they were raised against. We tested the specificity for each polyclonal serum using FFPE tissue sections known to be infected with T. gondii and N. caninum. The anti–Neospora-rNcSRS2 serum labeled specifically only N. caninum–infected tissue blocks, and the anti–Toxoplasma-rTgSRS2 serum was specific to only T. gondii–infected tissues. Moreover, tissues from 52 cattle and 19 sheep previously diagnosed by lesion profiles were tested using IHC with our polyclonal sera and PCR. The overall agreement between IHC and PCR was 90.1% for both polyclonal anti-rNcSRS2 and anti-rTgSRS2 sera. The polyclonal antisera were specific and allowed visual confirmation of protozoan parasites by IHC, but they were not as sensitive as PCR testing.EEA BalcarceFil: Lepore, Tanja. Moredun Research Institute; Reino UnidoFil: Macrae, Alastair I. Royal School of Veterinary Studies; Reino UnidoFil: Cantón, Germán. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; ArgentinaFil: Cantile, Carlo. Department of Veterinary Sciences; ItaliaFil: Martineau, Henny M. Royal Veterinary College. Department of Infectious Diseases; Reino UnidoFil: Palarea-Albaladejo, Javier. Biomathematics and Statistics Scotland; Reino UnidoFil: Cahalan, Stephen. Royal Veterinary College. Department of Infectious Diseases; Reino UnidoFil: Underwood, Clare. Moredun Research Institute; Reino UnidoFil: Katzer, Frank. Moredun Research Institute; Reino UnidoFil: Chianin, Francesca. Moredun Research Institute; Reino UnidoSage Publications2026-08-19T11:41:02Z2026-08-19T11:41:02Z2024-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/27463https://journals.sagepub.com/doi/10.1177/104063872412343221943-4936https://doi.org/10.1177/10406387241234Journal of Veterinary Diagnostic Investigation 36 (3) : 418-427 (2024)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/restrictedAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2026-09-24T11:43:43Zoai:localhost:20.500.12123/27463instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2026-09-24 11:43:44.077INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Evaluation of species-specific polyclonal antibodies to detect and differentiate between Neospora caninum and Toxoplasma gondii
title Evaluation of species-specific polyclonal antibodies to detect and differentiate between Neospora caninum and Toxoplasma gondii
spellingShingle Evaluation of species-specific polyclonal antibodies to detect and differentiate between Neospora caninum and Toxoplasma gondii
Lepore, Tanja
Neospora caninum
Toxoplasma gondii
Neosporosi
Toxoplasmosis
Aborto
Ganado
Anticuerpos Policlonales
Neosporosis
Abortion
Livestock
Polyclonal Antibodies
title_short Evaluation of species-specific polyclonal antibodies to detect and differentiate between Neospora caninum and Toxoplasma gondii
title_full Evaluation of species-specific polyclonal antibodies to detect and differentiate between Neospora caninum and Toxoplasma gondii
title_fullStr Evaluation of species-specific polyclonal antibodies to detect and differentiate between Neospora caninum and Toxoplasma gondii
title_full_unstemmed Evaluation of species-specific polyclonal antibodies to detect and differentiate between Neospora caninum and Toxoplasma gondii
title_sort Evaluation of species-specific polyclonal antibodies to detect and differentiate between Neospora caninum and Toxoplasma gondii
dc.creator.none.fl_str_mv Lepore, Tanja
Macrae, Alastair I.
Canton, German Jose
Cantile, Carlo
Martineau, Henny M.
Palarea-Albaladejo, Javier
Cahalan, Stephen
Underwood, Clare
Katzer, Frank
Chianini, Francesca
author Lepore, Tanja
author_facet Lepore, Tanja
Macrae, Alastair I.
Canton, German Jose
Cantile, Carlo
Martineau, Henny M.
Palarea-Albaladejo, Javier
Cahalan, Stephen
Underwood, Clare
Katzer, Frank
Chianini, Francesca
author_role author
author2 Macrae, Alastair I.
Canton, German Jose
Cantile, Carlo
Martineau, Henny M.
Palarea-Albaladejo, Javier
Cahalan, Stephen
Underwood, Clare
Katzer, Frank
Chianini, Francesca
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Neospora caninum
Toxoplasma gondii
Neosporosi
Toxoplasmosis
Aborto
Ganado
Anticuerpos Policlonales
Neosporosis
Abortion
Livestock
Polyclonal Antibodies
topic Neospora caninum
Toxoplasma gondii
Neosporosi
Toxoplasmosis
Aborto
Ganado
Anticuerpos Policlonales
Neosporosis
Abortion
Livestock
Polyclonal Antibodies
dc.description.none.fl_txt_mv Neosporosis and toxoplasmosis are major causes of abortion in livestock worldwide, leading to substantial economic losses. Detection tools are fundamental to the diagnosis and management of those diseases. Current immunohistochemistry (IHC) tests, using sera raised against whole parasite lysates, have not been able to distinguish between Toxoplasma gondii and Neospora caninum. We used T. gondii and N. caninum recombinant proteins, expressed in Escherichia coli and purified using insoluble conditions, to produce specific polyclonal rabbit antisera. We aimed to develop species-specific sera that could be used in IHC on formalin-fixed, paraffin-embedded (FFPE) tissue sections to improve the diagnosis of ruminant abortions caused by protozoa. Two polyclonal rabbit sera, raised against recombinant proteins, anti–Neospora-rNcSRS2 and anti–Toxoplasma-rTgSRS2, had specificity for the parasite they were raised against. We tested the specificity for each polyclonal serum using FFPE tissue sections known to be infected with T. gondii and N. caninum. The anti–Neospora-rNcSRS2 serum labeled specifically only N. caninum–infected tissue blocks, and the anti–Toxoplasma-rTgSRS2 serum was specific to only T. gondii–infected tissues. Moreover, tissues from 52 cattle and 19 sheep previously diagnosed by lesion profiles were tested using IHC with our polyclonal sera and PCR. The overall agreement between IHC and PCR was 90.1% for both polyclonal anti-rNcSRS2 and anti-rTgSRS2 sera. The polyclonal antisera were specific and allowed visual confirmation of protozoan parasites by IHC, but they were not as sensitive as PCR testing.
EEA Balcarce
Fil: Lepore, Tanja. Moredun Research Institute; Reino Unido
Fil: Macrae, Alastair I. Royal School of Veterinary Studies; Reino Unido
Fil: Cantón, Germán. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina
Fil: Cantile, Carlo. Department of Veterinary Sciences; Italia
Fil: Martineau, Henny M. Royal Veterinary College. Department of Infectious Diseases; Reino Unido
Fil: Palarea-Albaladejo, Javier. Biomathematics and Statistics Scotland; Reino Unido
Fil: Cahalan, Stephen. Royal Veterinary College. Department of Infectious Diseases; Reino Unido
Fil: Underwood, Clare. Moredun Research Institute; Reino Unido
Fil: Katzer, Frank. Moredun Research Institute; Reino Unido
Fil: Chianin, Francesca. Moredun Research Institute; Reino Unido
description Neosporosis and toxoplasmosis are major causes of abortion in livestock worldwide, leading to substantial economic losses. Detection tools are fundamental to the diagnosis and management of those diseases. Current immunohistochemistry (IHC) tests, using sera raised against whole parasite lysates, have not been able to distinguish between Toxoplasma gondii and Neospora caninum. We used T. gondii and N. caninum recombinant proteins, expressed in Escherichia coli and purified using insoluble conditions, to produce specific polyclonal rabbit antisera. We aimed to develop species-specific sera that could be used in IHC on formalin-fixed, paraffin-embedded (FFPE) tissue sections to improve the diagnosis of ruminant abortions caused by protozoa. Two polyclonal rabbit sera, raised against recombinant proteins, anti–Neospora-rNcSRS2 and anti–Toxoplasma-rTgSRS2, had specificity for the parasite they were raised against. We tested the specificity for each polyclonal serum using FFPE tissue sections known to be infected with T. gondii and N. caninum. The anti–Neospora-rNcSRS2 serum labeled specifically only N. caninum–infected tissue blocks, and the anti–Toxoplasma-rTgSRS2 serum was specific to only T. gondii–infected tissues. Moreover, tissues from 52 cattle and 19 sheep previously diagnosed by lesion profiles were tested using IHC with our polyclonal sera and PCR. The overall agreement between IHC and PCR was 90.1% for both polyclonal anti-rNcSRS2 and anti-rTgSRS2 sera. The polyclonal antisera were specific and allowed visual confirmation of protozoan parasites by IHC, but they were not as sensitive as PCR testing.
publishDate 2024
dc.date.none.fl_str_mv 2024-02
2026-08-19T11:41:02Z
2026-08-19T11:41:02Z
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12123/27463
https://journals.sagepub.com/doi/10.1177/10406387241234322
1943-4936
https://doi.org/10.1177/10406387241234
url http://hdl.handle.net/20.500.12123/27463
https://journals.sagepub.com/doi/10.1177/10406387241234322
https://doi.org/10.1177/10406387241234
identifier_str_mv 1943-4936
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv 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)
eu_rights_str_mv restrictedAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Sage Publications
publisher.none.fl_str_mv Sage Publications
dc.source.none.fl_str_mv Journal of Veterinary Diagnostic Investigation 36 (3) : 418-427 (2024)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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