RESUMO
Nucleotide sequence and phylogenetic analysis of the VP1 structural protein have been used extensively as diagnostic and epidemiological tools for foot and mouth disease virus (FMDV). In this report we have applied this methodology to the analysis of the VP1 coding sequence from FMDV strains isolated in Argentina during 1993-1994. The results demonstrated that the field isolates were related to the vaccine strains used at that time. However the involvement of the vaccine virus appeared to be different for outbreaks caused by FMD viruses type O or C. These data provide a database essential for determining the origin of new epizootics.
Assuntos
Animais , Bovinos , Aphthovirus , Doenças dos Bovinos/virologia , Febre Aftosa , Antígenos Virais/genética , Antígenos Virais/imunologia , Aphthovirus , Argentina , Sequência de Bases , Proteínas do Capsídeo , Surtos de Doenças , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/transmissão , Febre Aftosa , Dados de Sequência Molecular , Filogenia , Estudos Retrospectivos , RNA Viral , Alinhamento de Sequência , Homologia de Sequência do Ácido Nucleico , Sorotipagem , Vacinas ViraisRESUMO
The BVDV glycoproteins gp48 and gp53 were expressed in the baculovirus eukaryotic system. Both recombinant proteins were recognized in western blot analysis by monoclonal antibodies and polyclonal serum. Immunofluorescent test demonstrated that gp53 was localized on the cell surface whereas gp48 was in the cytoplasm. The expressed proteins were extracted by non-denaturing detergent treatment. Rabbit antiserum raised against gp53 recombinant protein efficiently neutralized the virus. These results demonstrate that the recombinant proteins have immunological properties similar to those of the native viral protein and that they can be useful as diagnostic reagents.
Assuntos
Animais , Bovinos , Masculino , Coelhos , Proteínas do Envelope Viral/isolamento & purificação , Vírus da Diarreia Viral Bovina/química , Western Blotting , Linhagem Celular , Soros Imunes , Rim , Nucleopoliedrovírus , Proteínas do Envelope Viral/genética , Proteínas do Envelope Viral/imunologia , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/isolamento & purificação , Spodoptera , Testículo/citologia , Transfecção , Vetores Genéticos/genética , Vírus da Diarreia Viral Bovina/genética , Vírus da Diarreia Viral Bovina/imunologiaRESUMO
Kinetics of the humoral immune primary response and seven-day secondary response of adult CF1 mice to FMDV O1 Campos adjuvanted in aluminium hydroxide-saponin (AHS) or in oil emulsion (OE) were evaluated by means of ELISA and passive hemagglutination (PH). Analysis of the response to AHS vaccine showed that ELISA measured maximal titres of primary response at 23 days post-vaccination (dpv), and at day 17 of secondary response, while PH detected maximal titres for primary as well as secondary response around day 60 pv. Mice immunized with OE vaccine studied by ELISA presented a plateau of primary response around 40 dpv, while secondary response was maximal around 80 dpv. The same sera tested by PH showed the highest titres for primary response on day 50 pv and secondary response was maximal on day 40 pv. A criterion for the evaluation of vaccination efficiency in the murine model is proposed based on the methods employed in the determination of antibody level.
RESUMO
Kinetics of the humoral immune primary response and seven-day secondary response of adult CF1 mice to FMDV O1 Campos adjuvanted in aluminium hydroxide-saponin (AHS) or in oil emulsion (OE) were evaluated by means of ELISA and passive hemagglutination (PH). Analysis of the response to AHS vaccine showed that ELISA measured maximal titres of primary response at 23 days post-vaccination (dpv), and at day 17 of secondary response, while PH detected maximal titres for primary as well as secondary response around day 60 pv. Mice immunized with OE vaccine studied by ELISA presented a plateau of primary response around 40 dpv, while secondary response was maximal around 80 dpv. The same sera tested by PH showed the highest titres for primary response on day 50 pv and secondary response was maximal on day 40 pv. A criterion for the evaluation of vaccination efficiency in the murine model is proposed based on the methods employed in the determination of antibody level.