RESUMEN
The goal of this project was to develop an alternative foot and mouth disease (FMD) vaccine candidate based on a recombinant protein consisting of efficient viral epitopes. A recombinant gene was designed that encodes B-cell epitopes of proteins VP1 and VP4 and T-cell epitopes of proteins 2C and 3D. The polyepitope protein (H-PE) was produced in E. coli bacteria or in N. benthamiana plants using a phytovirus expression system. The methods of extraction and purification of H-PE proteins from bacteria and plants were developed. Immunization of guinea pigs with the purified H-PE proteins induced an efficient immune response against foot and mouth disease virus (FMDV) serotype O/Taiwan/99 and protection against the disease. The polyepitope protein H-PE can be used as a basis for developing a new recombinant vaccine against FMD.
Asunto(s)
Escherichia coli/genética , Virus de la Fiebre Aftosa/inmunología , Nicotiana/genética , Proteínas Virales/biosíntesis , Proteínas Virales/inmunología , Vacunas Virales/biosíntesis , Vacunas Virales/inmunología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Epítopos/inmunología , Escherichia coli/citología , Cobayas , Datos de Secuencia Molecular , Nicotiana/citología , Vacunas Sintéticas/biosíntesis , Vacunas Sintéticas/química , Vacunas Sintéticas/inmunología , Vacunas Sintéticas/aislamiento & purificación , Proteínas Virales/química , Proteínas Virales/aislamiento & purificación , Vacunas Virales/química , Vacunas Virales/aislamiento & purificaciónRESUMEN
The study has shown the efficiency of a competitive ELISA (C-ELISA) variant or an indirect ELISA (I-ELUSA) in the detection of antibodies to swine vesicular disease virus (SVDV) versus traditional assays, such as a microneutralization test, a blocking ELIDA test, and a the reference test Ceditest SVDV (Cedi-Diagnostics B.V., Netherlands). Specific antibodies in the pig sera could be detected by C-ELISA on days 4-5 and by I-ELISA on day 6 after experimental SVDV infection. Specific antibodies were detected in a contact pig 11 days after the beginning of the experiment.