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.
Asunto(s)
Anticuerpos Antivirales/sangre , Enterovirus Humano B/aislamiento & purificación , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente Directa , Técnica del Anticuerpo Fluorescente Indirecta , Enfermedad Vesicular Porcina/diagnóstico , Animales , Enterovirus Humano B/inmunología , Porcinos , Enfermedad Vesicular Porcina/sangreRESUMEN
An indirect ELISA (I-ELISA) has been developed for swine vesicular disease virus-specific antibody detection. The analytic sensitivity of I-ELISA testing of serum samples from experimentally infected pigs with the known VN titer was 2 log2. Its diagnostic specificity was demonstrated as 100% in 4485 swine serum samples from different regions of the Russian Federation.
Asunto(s)
Anticuerpos Antivirales/sangre , Enterovirus Humano B/aislamiento & purificación , Ensayo de Inmunoadsorción Enzimática/veterinaria , Enfermedad Vesicular Porcina/diagnóstico , Animales , Anticuerpos Antivirales/inmunología , Especificidad de Anticuerpos , Antígenos Virales/inmunología , Enterovirus Humano B/inmunología , Sensibilidad y Especificidad , PorcinosRESUMEN
A field chick infectious bronchitis virus (IBV) was isolated from the pathological material on chick embryos. The nucleotide sequence of the S1 gene was determined and comparatively analyzed with some sequences of this gene of foreign and Russian vaccine strains and isolates. A cross-neutralization test using sera to various IBV seroptypes was performed. The isolate was shown to antigenically differ from the reference strains. Bioassay was carried out, by using one-day chicks and the immunogenic properties of the virus were investigated.