Diseño y desarrollo de un sistema inmunocromatográfico para la detección del virus de la diarrea viral bovina.
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Date
2025
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Publisher
Universidad de Concepción
Abstract
El virus de la diarrea viral bovina es un problema sanitario en los planteles bovinos alrededor del mundo. Actualmente, con el desarrollo de los métodos de diagnósticos, ha sido posible la implementación de programas de control y erradicación para este patógeno basados en la detección de animales persistentemente infectados (PI). En nuestro país, según el Servicio Agrícola y Ganadero, la infección con este virus es de notificación obligatoria, pero no existe ningún programa para el control de esta enfermedad y tampoco dispositivos que permitan la evaluación de los animales en terreno. El objetivo de este proyecto es el diseño y generación de un sistema de detección rápida por inmunocromatografía (tira reactiva o prueba de flujo lateral) para el virus de la diarrea viral bovina. Para esto se produjo la proteína de la cápside viral Erns de manera recombinante en células de insecto, con la cual se realizó la inmunización de ratones y gallinas para la obtención de anticuerpos monoclonales IgG y aviares IgY, respectivamente. Los anticuerpos monoclonales obtenidos se marcaron con nanopartículas de oro (AuNPs), y los anticuerpos aviares IgY anti rErns e IgG anti-ratón se imprimieron en una membrana de nitrocelulosa para el montaje final del sistema inmunocromatográfico. El ensayo de flujo lateral propuesto detecta hasta 25 ng de la proteína rErns demostrando que puede convertirse en una herramienta para el diagnóstico de animales PI con el VDVB.
Bovine viral diarrhea virus is a health problem in cattle herds around the world. Currently, with the development of diagnostic methods, it has been possible to implement control and eradication programs for this pathogen based on the detection of persistently infected animals (PI). In our country, according to the Servicio Agrícola y Ganadero, infection with this virus is notifiable, but there is no program for the control of this disease and there are no devices that allow the evaluation of animals in the field. The aim of this project is to design and generate a rapid detection system by immunochromatography (test strip or lateral flow test) for bovine viral diarrhea virus. For this purpose, the viral capsid protein Erns was produced recombinantly in insect cells, with which mice and hens were immunized to obtain IgG monoclonal antibodies and IgY antibodies, respectively. The monoclonal antibodies were labelled with gold nanoparticles (AuNPs), and the avian IgY anti-Erns and IgG anti-mouse antibodies were printed on a nitrocellulose membrane for the final assembly of the immunochromatographic system. The proposed lateral flow assay detects up to 25 ng of rErns protein demonstrating that it can become a tool for the diagnosis of PI animals with BVDV.
Bovine viral diarrhea virus is a health problem in cattle herds around the world. Currently, with the development of diagnostic methods, it has been possible to implement control and eradication programs for this pathogen based on the detection of persistently infected animals (PI). In our country, according to the Servicio Agrícola y Ganadero, infection with this virus is notifiable, but there is no program for the control of this disease and there are no devices that allow the evaluation of animals in the field. The aim of this project is to design and generate a rapid detection system by immunochromatography (test strip or lateral flow test) for bovine viral diarrhea virus. For this purpose, the viral capsid protein Erns was produced recombinantly in insect cells, with which mice and hens were immunized to obtain IgG monoclonal antibodies and IgY antibodies, respectively. The monoclonal antibodies were labelled with gold nanoparticles (AuNPs), and the avian IgY anti-Erns and IgG anti-mouse antibodies were printed on a nitrocellulose membrane for the final assembly of the immunochromatographic system. The proposed lateral flow assay detects up to 25 ng of rErns protein demonstrating that it can become a tool for the diagnosis of PI animals with BVDV.
Description
Tesis presentada para optar al grado de Doctor en Biotecnología Molecular.
Keywords
Tests inmunológicos, Virología veterinaria, Enfermedades transmisibles