Validation of an in house iELISA for serodiagnosis of caprine brucellosis and evaluation of the performance of a B. neotomae lysate for the detection of anti-smooth Brucella specif...

Autores
Foster, Camila Nayla; Rossi, Ursula Amaranta; Rossetti, Carlos Alberto
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Enzyme-linked immunosorbent assay (ELISA) is a widely used and effective tool for detection of anti-Brucella antibodies in serum, easy to perform with high sensitivity and specificity. In this study, we validated an in-house indirect ELISA using B. melitensis whole cell lysate as antigen (Bm-WCL iELISA) for the serodiagnosis of caprine brucellosis and evaluated the use of BSL-2 B. neotomae in replacement of BSL-3 Brucella species as an antigen for the detection of Brucella-specific antibodies in ruminant sera. Using 724 serum samples from female crossbred goats classified as brucellosis-positive or -negative by both the buffered plate antigen (BPA) and the complement fixation (CF) tests, the Bm-WCL iELISA was successfully validated with a sensitivity (Se) of 91.83 % (88.51–94.25 %) and a specificity (Sp) of 97.41 % (95.41–98.70 %). In addition, the Bm-WCL iELISA showed a great concordance with a commercial iELISA kit (k = 0.94) in a subset of 217 serum samples. To avoid working with a BSL-3 Brucella for antigen preparation, we replaced it with a less virulent Brucella species such as B. neotomae. A total of 214 goat and 220 cow serum samples were evaluated for the diagnosis of brucellosis using the B. neotomae whole cell homogenate (Bn-WCL) iELISA. The analysis of the ROC curves suggested cut-off values of 63.83 PP for goats and 24.04 PP for cattle, with associated Se and Sp of 98.18 % (93.61–99.68 %) and 90.38 % (83.20–94.69 %) respectively in goat sera, and 95.45 % (89.80–98.04 %) and 96.36 % (91.02–98.58 %) of Se and Sp, respectively in cattle. These results confirm the utility of the in house Bm-WCL iELISA and encourage validation of the Bn-WCL iELISA for the serodiagnosis of ruminant brucellosis in resource-limited areas where the disease is endemic.
Instituto de Patobiología
Fil: Foster, Camila Nayla. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; Argentina
Fil: Foster, Camila Nayla. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Rossi, Ursula Amaranta. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; Argentina
Fil: Rossi, Ursula Amaranta. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Rossetti, Carlos Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; Argentina
Fil: Rossetti, Carlos Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fuente
Veterinary Microbiology 302 : 110389. (March 2025)
Materia
Caprinae
Animal Diseases
Brucellosis
ELISA
Immunodiagnosis
Antibodies
Brucella
Enfermedades de los Animales
Brucelosis
Inmunodiagnóstico
Anticuerpos
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:20.500.12123/21368
Acceder
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oai_identifier_str oai:localhost:20.500.12123/21368
network_acronym_str INTADig
repository_id_str l
network_name_str INTA Digital (INTA)
spelling Validation of an in house iELISA for serodiagnosis of caprine brucellosis and evaluation of the performance of a B. neotomae lysate for the detection of anti-smooth Brucella specific antibodies in ruminantsFoster, Camila NaylaRossi, Ursula AmarantaRossetti, Carlos AlbertoCaprinaeAnimal DiseasesBrucellosisELISAImmunodiagnosisAntibodiesBrucellaEnfermedades de los AnimalesBrucelosisInmunodiagnósticoAnticuerposEnzyme-linked immunosorbent assay (ELISA) is a widely used and effective tool for detection of anti-Brucella antibodies in serum, easy to perform with high sensitivity and specificity. In this study, we validated an in-house indirect ELISA using B. melitensis whole cell lysate as antigen (Bm-WCL iELISA) for the serodiagnosis of caprine brucellosis and evaluated the use of BSL-2 B. neotomae in replacement of BSL-3 Brucella species as an antigen for the detection of Brucella-specific antibodies in ruminant sera. Using 724 serum samples from female crossbred goats classified as brucellosis-positive or -negative by both the buffered plate antigen (BPA) and the complement fixation (CF) tests, the Bm-WCL iELISA was successfully validated with a sensitivity (Se) of 91.83 % (88.51–94.25 %) and a specificity (Sp) of 97.41 % (95.41–98.70 %). In addition, the Bm-WCL iELISA showed a great concordance with a commercial iELISA kit (k = 0.94) in a subset of 217 serum samples. To avoid working with a BSL-3 Brucella for antigen preparation, we replaced it with a less virulent Brucella species such as B. neotomae. A total of 214 goat and 220 cow serum samples were evaluated for the diagnosis of brucellosis using the B. neotomae whole cell homogenate (Bn-WCL) iELISA. The analysis of the ROC curves suggested cut-off values of 63.83 PP for goats and 24.04 PP for cattle, with associated Se and Sp of 98.18 % (93.61–99.68 %) and 90.38 % (83.20–94.69 %) respectively in goat sera, and 95.45 % (89.80–98.04 %) and 96.36 % (91.02–98.58 %) of Se and Sp, respectively in cattle. These results confirm the utility of the in house Bm-WCL iELISA and encourage validation of the Bn-WCL iELISA for the serodiagnosis of ruminant brucellosis in resource-limited areas where the disease is endemic.Instituto de PatobiologíaFil: Foster, Camila Nayla. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; ArgentinaFil: Foster, Camila Nayla. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rossi, Ursula Amaranta. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; ArgentinaFil: Rossi, Ursula Amaranta. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rossetti, Carlos Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; ArgentinaFil: Rossetti, Carlos Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaElsevier2025-02-20T11:58:13Z2025-02-20T11:58:13Z2025-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/21368https://www.sciencedirect.com/science/article/abs/pii/S03781135250002400378-11351873-2542https://doi.org/10.1016/j.vetmic.2025.110389Veterinary Microbiology 302 : 110389. (March 2025)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repograntAgreement/INTA/2019-PD-E5-I103-001, Desarrollo de tecnologías diagnósticas y estudios epidemiológicos para el control de enfermedades que afectan la producción animal y la salud públicainfo: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)2025-09-04T09:50:55Zoai:localhost:20.500.12123/21368instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-04 09:50:56.291INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Validation of an in house iELISA for serodiagnosis of caprine brucellosis and evaluation of the performance of a B. neotomae lysate for the detection of anti-smooth Brucella specific antibodies in ruminants
title Validation of an in house iELISA for serodiagnosis of caprine brucellosis and evaluation of the performance of a B. neotomae lysate for the detection of anti-smooth Brucella specific antibodies in ruminants
spellingShingle Validation of an in house iELISA for serodiagnosis of caprine brucellosis and evaluation of the performance of a B. neotomae lysate for the detection of anti-smooth Brucella specific antibodies in ruminants
Foster, Camila Nayla
Caprinae
Animal Diseases
Brucellosis
ELISA
Immunodiagnosis
Antibodies
Brucella
Enfermedades de los Animales
Brucelosis
Inmunodiagnóstico
Anticuerpos
title_short Validation of an in house iELISA for serodiagnosis of caprine brucellosis and evaluation of the performance of a B. neotomae lysate for the detection of anti-smooth Brucella specific antibodies in ruminants
title_full Validation of an in house iELISA for serodiagnosis of caprine brucellosis and evaluation of the performance of a B. neotomae lysate for the detection of anti-smooth Brucella specific antibodies in ruminants
title_fullStr Validation of an in house iELISA for serodiagnosis of caprine brucellosis and evaluation of the performance of a B. neotomae lysate for the detection of anti-smooth Brucella specific antibodies in ruminants
title_full_unstemmed Validation of an in house iELISA for serodiagnosis of caprine brucellosis and evaluation of the performance of a B. neotomae lysate for the detection of anti-smooth Brucella specific antibodies in ruminants
title_sort Validation of an in house iELISA for serodiagnosis of caprine brucellosis and evaluation of the performance of a B. neotomae lysate for the detection of anti-smooth Brucella specific antibodies in ruminants
dc.creator.none.fl_str_mv Foster, Camila Nayla
Rossi, Ursula Amaranta
Rossetti, Carlos Alberto
author Foster, Camila Nayla
author_facet Foster, Camila Nayla
Rossi, Ursula Amaranta
Rossetti, Carlos Alberto
author_role author
author2 Rossi, Ursula Amaranta
Rossetti, Carlos Alberto
author2_role author
author
dc.subject.none.fl_str_mv Caprinae
Animal Diseases
Brucellosis
ELISA
Immunodiagnosis
Antibodies
Brucella
Enfermedades de los Animales
Brucelosis
Inmunodiagnóstico
Anticuerpos
topic Caprinae
Animal Diseases
Brucellosis
ELISA
Immunodiagnosis
Antibodies
Brucella
Enfermedades de los Animales
Brucelosis
Inmunodiagnóstico
Anticuerpos
dc.description.none.fl_txt_mv Enzyme-linked immunosorbent assay (ELISA) is a widely used and effective tool for detection of anti-Brucella antibodies in serum, easy to perform with high sensitivity and specificity. In this study, we validated an in-house indirect ELISA using B. melitensis whole cell lysate as antigen (Bm-WCL iELISA) for the serodiagnosis of caprine brucellosis and evaluated the use of BSL-2 B. neotomae in replacement of BSL-3 Brucella species as an antigen for the detection of Brucella-specific antibodies in ruminant sera. Using 724 serum samples from female crossbred goats classified as brucellosis-positive or -negative by both the buffered plate antigen (BPA) and the complement fixation (CF) tests, the Bm-WCL iELISA was successfully validated with a sensitivity (Se) of 91.83 % (88.51–94.25 %) and a specificity (Sp) of 97.41 % (95.41–98.70 %). In addition, the Bm-WCL iELISA showed a great concordance with a commercial iELISA kit (k = 0.94) in a subset of 217 serum samples. To avoid working with a BSL-3 Brucella for antigen preparation, we replaced it with a less virulent Brucella species such as B. neotomae. A total of 214 goat and 220 cow serum samples were evaluated for the diagnosis of brucellosis using the B. neotomae whole cell homogenate (Bn-WCL) iELISA. The analysis of the ROC curves suggested cut-off values of 63.83 PP for goats and 24.04 PP for cattle, with associated Se and Sp of 98.18 % (93.61–99.68 %) and 90.38 % (83.20–94.69 %) respectively in goat sera, and 95.45 % (89.80–98.04 %) and 96.36 % (91.02–98.58 %) of Se and Sp, respectively in cattle. These results confirm the utility of the in house Bm-WCL iELISA and encourage validation of the Bn-WCL iELISA for the serodiagnosis of ruminant brucellosis in resource-limited areas where the disease is endemic.
Instituto de Patobiología
Fil: Foster, Camila Nayla. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; Argentina
Fil: Foster, Camila Nayla. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Rossi, Ursula Amaranta. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; Argentina
Fil: Rossi, Ursula Amaranta. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Rossetti, Carlos Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria; Argentina
Fil: Rossetti, Carlos Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description Enzyme-linked immunosorbent assay (ELISA) is a widely used and effective tool for detection of anti-Brucella antibodies in serum, easy to perform with high sensitivity and specificity. In this study, we validated an in-house indirect ELISA using B. melitensis whole cell lysate as antigen (Bm-WCL iELISA) for the serodiagnosis of caprine brucellosis and evaluated the use of BSL-2 B. neotomae in replacement of BSL-3 Brucella species as an antigen for the detection of Brucella-specific antibodies in ruminant sera. Using 724 serum samples from female crossbred goats classified as brucellosis-positive or -negative by both the buffered plate antigen (BPA) and the complement fixation (CF) tests, the Bm-WCL iELISA was successfully validated with a sensitivity (Se) of 91.83 % (88.51–94.25 %) and a specificity (Sp) of 97.41 % (95.41–98.70 %). In addition, the Bm-WCL iELISA showed a great concordance with a commercial iELISA kit (k = 0.94) in a subset of 217 serum samples. To avoid working with a BSL-3 Brucella for antigen preparation, we replaced it with a less virulent Brucella species such as B. neotomae. A total of 214 goat and 220 cow serum samples were evaluated for the diagnosis of brucellosis using the B. neotomae whole cell homogenate (Bn-WCL) iELISA. The analysis of the ROC curves suggested cut-off values of 63.83 PP for goats and 24.04 PP for cattle, with associated Se and Sp of 98.18 % (93.61–99.68 %) and 90.38 % (83.20–94.69 %) respectively in goat sera, and 95.45 % (89.80–98.04 %) and 96.36 % (91.02–98.58 %) of Se and Sp, respectively in cattle. These results confirm the utility of the in house Bm-WCL iELISA and encourage validation of the Bn-WCL iELISA for the serodiagnosis of ruminant brucellosis in resource-limited areas where the disease is endemic.
publishDate 2025
dc.date.none.fl_str_mv 2025-02-20T11:58:13Z
2025-02-20T11:58:13Z
2025-03
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/20.500.12123/21368
https://www.sciencedirect.com/science/article/abs/pii/S0378113525000240
0378-1135
1873-2542
https://doi.org/10.1016/j.vetmic.2025.110389
url http://hdl.handle.net/20.500.12123/21368
https://www.sciencedirect.com/science/article/abs/pii/S0378113525000240
https://doi.org/10.1016/j.vetmic.2025.110389
identifier_str_mv 0378-1135
1873-2542
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repograntAgreement/INTA/2019-PD-E5-I103-001, Desarrollo de tecnologías diagnósticas y estudios epidemiológicos para el control de enfermedades que afectan la producción animal y la salud pública
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 Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv Veterinary Microbiology 302 : 110389. (March 2025)
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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