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Intervirology ; 56(1): 55-9, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23052035

RESUMEN

AIMS: To develop an effective diagnostic kit, based on a competitive ELISA-based system (cELISA), for detecting serum antibody against peste des petits ruminants virus (PPRV). METHODS: Epitope peptides of the nucleocapsid (N) protein of Tibetan PPRV were synthesized chemically and injected into rabbits to prepare hyperimmune antisera. Test sera were incubated simultaneously with hyperimmune antisera and added to the wells of ELISA plates coated previously with recombinant N protein. Horseradish peroxidase-conjugated goat anti-rabbit antibody was employed to detect the quantity of hyperimmune antisera combined with recombinant N protein. RESULTS: A cELISA has been developed for monitoring PPRV infections with a cutoff value of 35. Relative sensitivity and specificity values of the epitope-based cELISA were 96.18 and 91.29%, respectively, when compared with a commercial cELISA kit in a test involving 1,039 serum samples. CONCLUSION: We report an efficient method for preparing antibody suitable for incorporation into a cELISA that can be used routinely for the detection of PPRV antibodies in serum samples. The method eliminated the requirement for virus culture and monoclonal antibody preparation, reduced the biorisk posed by virus-dependent manipulations, and the performance of the resultant cELISA compared favorably with a commercially available cELISA kit.


Asunto(s)
Anticuerpos Antivirales/sangre , Ensayo de Inmunoadsorción Enzimática/veterinaria , Enfermedades de las Cabras/diagnóstico , Proteínas de la Nucleocápside/inmunología , Peste de los Pequeños Rumiantes/veterinaria , Virus de la Peste de los Pequeños Rumiantes/inmunología , Enfermedades de las Ovejas/diagnóstico , Animales , Ensayo de Inmunoadsorción Enzimática/métodos , Enfermedades de las Cabras/inmunología , Enfermedades de las Cabras/virología , Cabras , Peste de los Pequeños Rumiantes/diagnóstico , Peste de los Pequeños Rumiantes/inmunología , Conejos , Ovinos , Enfermedades de las Ovejas/inmunología , Enfermedades de las Ovejas/virología
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