Sensibilidad de una prueba de ELISA-indirecta para el diagnóstico de la fascioliasis animal utilizando antígenos de excreción-secreción de Fasciola hepatica
No Thumbnail Available
Date
2023
Journal Title
Journal ISSN
Volume Title
Publisher
Universidad Nacional de Trujillo
Abstract
La fascioliasis, zoonosis parasitaria de distribución mundial causada por Fasciola hepatica, es de importancia médico-veterinaria debido a que disminuye la producción pecuaria. Nuevos datos epidemiológicos sugieren que la fascioliasis está en plena expansión, por lo que es necesario contar con herramientas de diagnóstico útiles, que complementen al coprológico que es insatisfactorio por su baja sensibilidad. El objetivo de la presente investigación fue determinar la sensibilidad de una técnica de ELISA-indirecta utilizando antígenos de excreción- secreción (Ag.E-S) de las formas adultas de F. hepatica obtenidos en Medio Mínimo Esencial (MEM) a las 8 h de incubación en la detección de anticuerpos en suero de Bos taurus y Ovis aries infectados naturalmente y sacrificados en el Camal El Porvenir (Trujillo, Perú). Los Ag.E- S fueron obtenidos incubando ejemplares adultos vivos del parásito en MEM (bioRad). La capacidad antigénica se evaluó por el método de inmunoprecipitación en lámina y la concentración de proteínas por la técnica de Bradford. La concentración de proteínas fue de 124 μg/mL y el punto de corte (Cut-off, DO490) de 0.33 nm. La sensibilidad encontrada fue del 100
% cuando se evaluó los sueros de B. taurus (50/50) y 100 % cuando se evaluó los sueros de O. aries (50/50). En conclusión, la prueba de ELISA-indirecta evaluada con Ag.E-S de formas adultas de F. hepatica obtenidos a las 8 h de incubación en medio MEM, presenta una sensibilidad del 100 % en la detección de anticuerpos en los sueros de ovinos y vacunos infectados naturalmente; por lo tanto, podría ser empleada en estudios de seroprevalencia.
Fascioliasis, a parasitic zoonosis with worldwide distribution caused by Fasciola hepatica, is of medical-veterinary importance because it decreases livestock production. New epidemiological data suggest that fasciolosis is in full expansion, so it is necessary to have useful diagnostic tools that complement the stool test, which is unsatisfactory due to its low sensitivity. The objective of the present investigation was to determine the sensitivity of an indirect-ELISA technique using excretion-secretion antigens (Ag.E-S) of the adult forms of F. hepatica obtained in Minimum Essential Medium (MEM) at 8 h of incubation in the detection of antibodies in serum of Bos taurus and Ovis aries naturally infected and sacrificed in Camal El Porvenir (Trujillo, Peru). Ag.E-S was obtained by incubating live adult specimens of the parasite in MEM (bioRad). The antigenic capacity was evaluated by the slide immunoprecipitation method and the protein concentration by the Bradford technique. The protein concentration was 124 μg/mL and the cut-off point (Cut-off, OD490) was 0.33 nm. The sensitivity found was 100 % when evaluating the B. taurus sera (50/50) and 100 % when evaluating the O. aries sera (50/50). In conclusion, the indirect-ELISA test evaluated with Ag.E-S of adult forms of F. hepatica obtained after 8 h of incubation in MEM medium, presents a sensitivity of 100 % in the detection of antibodies in sera of naturally infected sheep and cattle; therefore, it could be used in seroprevalence studies.
Fascioliasis, a parasitic zoonosis with worldwide distribution caused by Fasciola hepatica, is of medical-veterinary importance because it decreases livestock production. New epidemiological data suggest that fasciolosis is in full expansion, so it is necessary to have useful diagnostic tools that complement the stool test, which is unsatisfactory due to its low sensitivity. The objective of the present investigation was to determine the sensitivity of an indirect-ELISA technique using excretion-secretion antigens (Ag.E-S) of the adult forms of F. hepatica obtained in Minimum Essential Medium (MEM) at 8 h of incubation in the detection of antibodies in serum of Bos taurus and Ovis aries naturally infected and sacrificed in Camal El Porvenir (Trujillo, Peru). Ag.E-S was obtained by incubating live adult specimens of the parasite in MEM (bioRad). The antigenic capacity was evaluated by the slide immunoprecipitation method and the protein concentration by the Bradford technique. The protein concentration was 124 μg/mL and the cut-off point (Cut-off, OD490) was 0.33 nm. The sensitivity found was 100 % when evaluating the B. taurus sera (50/50) and 100 % when evaluating the O. aries sera (50/50). In conclusion, the indirect-ELISA test evaluated with Ag.E-S of adult forms of F. hepatica obtained after 8 h of incubation in MEM medium, presents a sensitivity of 100 % in the detection of antibodies in sera of naturally infected sheep and cattle; therefore, it could be used in seroprevalence studies.
Description
Keywords
Antígenos de excreción-secreción, Bos taurus, ELISA-indirecta, Fasciola hepatica, Medio Mínimo Esencial, Ovis aries, sensibilidad