A standardized enzyme-linked immunosorbent assay (ELISA) for the detection of Infectious bronchitis virus antibodies in serum and tracheobronchial lavage samples of chickens
Infectious bronchitis (IB) is a major viral disease that causes substantial economic losses in the global poultry industry. Despite the implementation of extensive vaccination programs, it remains a persistent threat. Monitoring the antibody response against infectious bronchitis virus (IBV) is esse...
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| Formato: | Artículo |
| Lenguaje: | Inglés |
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American Association of Avian Pathologists
2025
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/24825 https://meridian.allenpress.com/avian-diseases/article/69/4/420/508188/A-Standardized-Enzyme-Linked-Immunosorbent-Assay https://doi.org/10.1637/aviandiseases-D-25-00050 |
| _version_ | 1855487404904808448 |
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| author | Di Giacomo, Sebastián Gerez Miranda, Rocio Del Carmen Olivera, Valeria Soledad Asenzo, Gustavo Daniel Jaton, Juan Marcelo Vagnozzi, Ariel Eduardo |
| author_browse | Asenzo, Gustavo Daniel Di Giacomo, Sebastián Gerez Miranda, Rocio Del Carmen Jaton, Juan Marcelo Olivera, Valeria Soledad Vagnozzi, Ariel Eduardo |
| author_facet | Di Giacomo, Sebastián Gerez Miranda, Rocio Del Carmen Olivera, Valeria Soledad Asenzo, Gustavo Daniel Jaton, Juan Marcelo Vagnozzi, Ariel Eduardo |
| author_sort | Di Giacomo, Sebastián |
| collection | INTA Digital |
| description | Infectious bronchitis (IB) is a major viral disease that causes substantial economic losses in the global poultry industry. Despite the implementation of extensive vaccination programs, it remains a persistent threat. Monitoring the antibody response against infectious bronchitis virus (IBV) is essential for studies focused on vaccine development, as well as for evaluating the effectiveness and proper administration of vaccines in poultry flocks. In this study we developed and optimized an indirect enzyme-linked immunosorbent assay (IB-ELISA) for the detection of IBV-specific antibodies. The use of purified virus, obtained through sucrose density gradient centrifugation, was critical for minimizing nonspecific background reactivity. A panel of antisera and tracheobronchial lavage samples from IBV-infected and inactivated-vaccine-immunized chickens was tested. The IB-ELISA demonstrated 100% sensitivity and specificity when compared with a commercial IBV ELISA kit used as the gold standard. The assay reliably detected IBV-specific IgG antibodies in both serum and tracheobronchial lavage samples, with positive responses observed as early as 7 days postinfection and 21 days postvaccination, respectively. Although the IB-ELISA could not differentiate between IBV serotypes because of cross-reactivity, it effectively identified antibodies against multiple strains. These findings indicate that the IB-ELISA is a valuable tool for assessing the effectiveness of vaccination programs and for studies requiring the evaluation of both mucosal and systemic IgG responses, including those involving experimental vaccines. |
| format | Artículo |
| id | INTA24825 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2025 |
| publishDateRange | 2025 |
| publishDateSort | 2025 |
| publisher | American Association of Avian Pathologists |
| publisherStr | American Association of Avian Pathologists |
| record_format | dspace |
| spelling | INTA248252025-12-30T15:17:14Z A standardized enzyme-linked immunosorbent assay (ELISA) for the detection of Infectious bronchitis virus antibodies in serum and tracheobronchial lavage samples of chickens Di Giacomo, Sebastián Gerez Miranda, Rocio Del Carmen Olivera, Valeria Soledad Asenzo, Gustavo Daniel Jaton, Juan Marcelo Vagnozzi, Ariel Eduardo ELISA Antibodies Chickens Bronchitis Infectious Diseases Immunoenzyme Techniques Anticuerpo Pollo Bronquitis Enfermedades Infecciosas Técnica Inmunoenzimática Serum Tracheobronchial Lavage Suero Lavaje Traqueobronquial Infectious bronchitis (IB) is a major viral disease that causes substantial economic losses in the global poultry industry. Despite the implementation of extensive vaccination programs, it remains a persistent threat. Monitoring the antibody response against infectious bronchitis virus (IBV) is essential for studies focused on vaccine development, as well as for evaluating the effectiveness and proper administration of vaccines in poultry flocks. In this study we developed and optimized an indirect enzyme-linked immunosorbent assay (IB-ELISA) for the detection of IBV-specific antibodies. The use of purified virus, obtained through sucrose density gradient centrifugation, was critical for minimizing nonspecific background reactivity. A panel of antisera and tracheobronchial lavage samples from IBV-infected and inactivated-vaccine-immunized chickens was tested. The IB-ELISA demonstrated 100% sensitivity and specificity when compared with a commercial IBV ELISA kit used as the gold standard. The assay reliably detected IBV-specific IgG antibodies in both serum and tracheobronchial lavage samples, with positive responses observed as early as 7 days postinfection and 21 days postvaccination, respectively. Although the IB-ELISA could not differentiate between IBV serotypes because of cross-reactivity, it effectively identified antibodies against multiple strains. These findings indicate that the IB-ELISA is a valuable tool for assessing the effectiveness of vaccination programs and for studies requiring the evaluation of both mucosal and systemic IgG responses, including those involving experimental vaccines. Nota de investigación- Un ensayo de inmunoabsorción con enzimas ligadas (ELISA) estandarizado para la detección de anticuerpos contra el virus de la bronquitis infecciosa en muestras de suero y lavado traqueobronquial de pollos. La bronquitis infecciosa es una enfermedad viral importante que causa pérdidas económicas sustanciales en la industria avícola a nivel mundial. A pesar de la implementación de extensos programas de vacunación, sigue siendo una amenaza persistente. El monitoreo de la respuesta de anticuerpos contra el virus de la bronquitis infecciosa es esencial para los estudios centrados en el desarrollo de vacunas, así como para evaluar la efectividad y la correcta administración de vacunas en parvadas avícolas. En este estudio, se desarrolló y optimizó un ensayo de inmunoabsorción con enzimas ligadas indirecto (IB-ELISA) para la detección de anticuerpos específicos contra el virus de la bronquitis infecciosa. El uso de un virus purificado, obtenido mediante centrifugación en gradiente de densidad de sacarosa, fue crucial para minimizar la reactividad de fondo inespecífica. Se analizó un panel de antisueros y muestras de lavado traqueobronquial de pollos infectados con el virus de la bronquitis infecciosa e inmunizados con una vacuna inactivada. La prueba IB-ELISA demostró una sensibilidad y especificidad del 100% en comparación con un kit comercial de ELISA para el virus de la bronquitis infecciosa, que fue utilizado como prueba estándar de oro. El ensayo detectó con confiabilidad anticuerpos IgG específicos contra el virus de la bronquitis infecciosa tanto en muestras de suero como de lavado traqueobronquial, con respuestas positivas observadas desde los 7 días posteriores a la infección hasta los 21 días posteriores a la vacunación. Si bien la prueba IB-ELISA no pudo diferenciar entre los serotipos del virus de la bronquitis infecciosa debido a la reactividad cruzada, identificó eficazmente anticuerpos contra múltiples cepas. Estos hallazgos indican que la prueba IB-ELISA es una herramienta valiosa para evaluar la eficacia de los programas de vacunación y para estudios que requieren la evaluación de las respuestas IgG tanto en las mucosas como sistémicas, incluyendo aquellos con vacunas experimentales. Instituto de Virología Fil: Di Giacomo, Sebastián. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina Fil: Di Giacomo, Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Gerez Miranda, Rocio Del Carmen. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina Fil: Gerez Miranda, Rocio Del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Olivera, Valeria Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina Fil: Olivera, Valeria Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Asenzo, Gustavo Daniel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina Fil: Asenzo, Gustavo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Jaton, Juan Marcelo. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Agrobiotecnología y Biología Molecular; Argentina Fil: Jaton, Juan Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Vagnozzi, Ariel Eduardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas; Argentina Fil: Vagnozzi, Ariel Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina 2025-12-30T15:09:30Z 2025-12-30T15:09:30Z 2025-12 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/24825 https://meridian.allenpress.com/avian-diseases/article/69/4/420/508188/A-Standardized-Enzyme-Linked-Immunosorbent-Assay 0005-2086 https://doi.org/10.1637/aviandiseases-D-25-00050 eng info:eu-repo/semantics/openAccess 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 American Association of Avian Pathologists Avian Diseases 69 (4) : 420-425 (December 2025) |
| spellingShingle | ELISA Antibodies Chickens Bronchitis Infectious Diseases Immunoenzyme Techniques Anticuerpo Pollo Bronquitis Enfermedades Infecciosas Técnica Inmunoenzimática Serum Tracheobronchial Lavage Suero Lavaje Traqueobronquial Di Giacomo, Sebastián Gerez Miranda, Rocio Del Carmen Olivera, Valeria Soledad Asenzo, Gustavo Daniel Jaton, Juan Marcelo Vagnozzi, Ariel Eduardo A standardized enzyme-linked immunosorbent assay (ELISA) for the detection of Infectious bronchitis virus antibodies in serum and tracheobronchial lavage samples of chickens |
| title | A standardized enzyme-linked immunosorbent assay (ELISA) for the detection of Infectious bronchitis virus antibodies in serum and tracheobronchial lavage samples of chickens |
| title_full | A standardized enzyme-linked immunosorbent assay (ELISA) for the detection of Infectious bronchitis virus antibodies in serum and tracheobronchial lavage samples of chickens |
| title_fullStr | A standardized enzyme-linked immunosorbent assay (ELISA) for the detection of Infectious bronchitis virus antibodies in serum and tracheobronchial lavage samples of chickens |
| title_full_unstemmed | A standardized enzyme-linked immunosorbent assay (ELISA) for the detection of Infectious bronchitis virus antibodies in serum and tracheobronchial lavage samples of chickens |
| title_short | A standardized enzyme-linked immunosorbent assay (ELISA) for the detection of Infectious bronchitis virus antibodies in serum and tracheobronchial lavage samples of chickens |
| title_sort | standardized enzyme linked immunosorbent assay elisa for the detection of infectious bronchitis virus antibodies in serum and tracheobronchial lavage samples of chickens |
| topic | ELISA Antibodies Chickens Bronchitis Infectious Diseases Immunoenzyme Techniques Anticuerpo Pollo Bronquitis Enfermedades Infecciosas Técnica Inmunoenzimática Serum Tracheobronchial Lavage Suero Lavaje Traqueobronquial |
| url | http://hdl.handle.net/20.500.12123/24825 https://meridian.allenpress.com/avian-diseases/article/69/4/420/508188/A-Standardized-Enzyme-Linked-Immunosorbent-Assay https://doi.org/10.1637/aviandiseases-D-25-00050 |
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