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
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 iEL...
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| Format: | info:ar-repo/semantics/artículo |
| Language: | Inglés |
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Elsevier
2025
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| Online Access: | 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 |
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| author | Foster, Camila Nayla Rossi, Ursula Amaranta Rossetti, Carlos Alberto |
| author_browse | Foster, Camila Nayla Rossetti, Carlos Alberto Rossi, Ursula Amaranta |
| author_facet | Foster, Camila Nayla Rossi, Ursula Amaranta Rossetti, Carlos Alberto |
| author_sort | Foster, Camila Nayla |
| collection | INTA Digital |
| 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. |
| format | info:ar-repo/semantics/artículo |
| id | INTA21368 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2025 |
| publishDateRange | 2025 |
| publishDateSort | 2025 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA213682025-02-20T12:03:34Z 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 Rossi, Ursula Amaranta Rossetti, Carlos Alberto Caprinae Animal Diseases Brucellosis ELISA Immunodiagnosis Antibodies Brucella Enfermedades de los Animales Brucelosis Inmunodiagnóstico Anticuerpos 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 2025-02-20T11:58:13Z 2025-02-20T11:58:13Z 2025-03 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 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 eng 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 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) application/pdf Elsevier Veterinary Microbiology 302 : 110389. (March 2025) |
| spellingShingle | Caprinae Animal Diseases Brucellosis ELISA Immunodiagnosis Antibodies Brucella Enfermedades de los Animales Brucelosis Inmunodiagnóstico Anticuerpos Foster, Camila Nayla Rossi, Ursula Amaranta Rossetti, Carlos Alberto 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 |
| 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_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_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 |
| topic | Caprinae Animal Diseases Brucellosis ELISA Immunodiagnosis Antibodies Brucella Enfermedades de los Animales Brucelosis Inmunodiagnóstico Anticuerpos |
| 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 |
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