RESUMO
During 2000-2002 a foot-and-mouth disease (FMD) epizootic affected Argentina and spread across the country resulting in more than 2500 outbreaks. In order to study the evolution of the FMD viruses (FMDV) and help with disease control measures, a genetic characterization and phylogenetic analysis was performed of 43 field isolates representative of the epizootic. The nucleotide sequence of the VP1-coding region was determined for the viruses and used in this study. Two serotype A lineages, A/Arg/00 and A/Arg/01 (1000/1000 bootstrap value) and two different serotype O/Arg/00 lineages (848/1000 bootstrap value) were identified. Phylogenetic analysis showed that viruses A/Arg/01 and O/Arg/00 could be related with former South American isolates, while the origin of A Argentina 2000 viruses remains unclear. Comparison of the amino acid sequences with vaccine reference strains revealed differences at critical antigenic sites for emergent strains A/Arg/00 and A/Arg/01, leading to a change in the current vaccine formulation.
Assuntos
Proteínas do Capsídeo/genética , Vírus da Febre Aftosa/genética , Febre Aftosa/epidemiologia , Sequência de Aminoácidos , Animais , Argentina/epidemiologia , Sequência de Bases , Proteínas do Capsídeo/química , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/virologia , Surtos de Doenças/veterinária , Febre Aftosa/virologia , Vírus da Febre Aftosa/classificação , Epidemiologia Molecular , Dados de Sequência Molecular , Filogenia , RNA Viral/química , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Alinhamento de Sequência/veterinária , Homologia de Sequência do Ácido Nucleico , Sorotipagem/veterináriaRESUMO
The complete nucleotide sequence of foot-and-mouth disease virus (FMDV) South American strain O(1) Campos/Bra/58 was determined. The 8,168 Kb sequence and the deduced amino acid sequence were compared to published FMDV sequences. They showed the highest sequence homology with the O(1) Kaufbeuren/FRG/66 strain, but closer evolutionary relatedness to the Argentinean strains.
Assuntos
Vírus da Febre Aftosa/genética , RNA Viral/química , Sequência de Aminoácidos , Sequência de Bases , Proteínas do Capsídeo/química , Vírus da Febre Aftosa/classificação , Dados de Sequência Molecular , Filogenia , Alinhamento de Sequência , Homologia de Sequência de AminoácidosRESUMO
We have analysed complete or partial VPI sequences of 31 foot-and-mouth disease (FMD) viruses belonging to serotypes A, O and C to determine the genetic relatedness of field strains of FMD virus (FMDV) that have circulated in Argentina between 1961 and 1994. Phylogenetic analysis, which also included 15 previously published Argentinean sequences and six reference strains, revealed that (i) FMD type A strains showed the highest genetic heterogeneity and could be divided into five lineages with a sequence divergence of 0.9-18.5% between strains (ii) most of the FMD type O viruses grouped in two clusters (within cluster sequence divergence ranging from 0.2% to 6.0%) circulating in Argentina since the early 1960s, and (iii) FMD type C viruses were grouped in two clusters with a 13.4% nucleotide sequence divergence between each cluster. The availability of sequence data for many more field isolates from the region will enable us to understand the genetic relationships between FMDV strains and to rapidly trace the source of an FMD outbreak for epidemiological surveillance.
Assuntos
Vírus da Febre Aftosa/classificação , Argentina/epidemiologia , Vírus da Febre Aftosa/genética , Epidemiologia Molecular , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
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
Aphthovirus/isolamento & purificação , Capsídeo/genética , Doenças dos Bovinos/virologia , Febre Aftosa/virologia , Animais , Antígenos Virais/genética , Antígenos Virais/imunologia , Aphthovirus/classificação , Aphthovirus/genética , Aphthovirus/imunologia , Argentina/epidemiologia , Sequência de Bases , Proteínas do Capsídeo , Bovinos , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/transmissão , Surtos de Doenças , Febre Aftosa/epidemiologia , Febre Aftosa/transmissão , Dados de Sequência Molecular , Filogenia , RNA Viral/genética , Estudos Retrospectivos , Alinhamento de Sequência , Homologia de Sequência do Ácido Nucleico , Sorotipagem , Vacinas Virais/efeitos adversosRESUMO
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
Vírus da Diarreia Viral Bovina/química , Proteínas do Envelope Viral/isolamento & purificação , Animais , Western Blotting , Bovinos , Linhagem Celular , Vírus da Diarreia Viral Bovina/genética , Vírus da Diarreia Viral Bovina/imunologia , Vetores Genéticos/genética , Soros Imunes , Rim/citologia , Masculino , Nucleopoliedrovírus/genética , Coelhos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/isolamento & purificação , Spodoptera/citologia , Testículo/citologia , Transfecção , Proteínas do Envelope Viral/genética , Proteínas do Envelope Viral/imunologiaRESUMO
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
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.(AU)
Assuntos
Estudo Comparativo , Animais , Bovinos , RESEARCH SUPPORT, NON-U.S. GOVT , Aphthovirus/isolamento & purificação , Doenças dos Bovinos/virologia , Febre Aftosa/virologia , Antígenos Virais/genética , Antígenos Virais/imunologia , Aphthovirus/classificação , Aphthovirus/genética , Aphthovirus/imunologia , Argentina/epidemiologia , Sequência de Bases , Proteínas do Capsídeo , Doenças dos Bovinos/epidemiologia , Doenças dos Bovinos/transmissão , Surtos de Doenças , Febre Aftosa/epidemiologia , Febre Aftosa/transmissão , Dados de Sequência Molecular , Filogenia , RNA Viral/genética , Estudos Retrospectivos , Alinhamento de Sequência , Homologia de Sequência do Ácido Nucleico , Sorotipagem , Vacinas Virais/efeitos adversosRESUMO
An analysis of the informative content of sequence stretches on the foot-and-mouth disease virus (FMDV) VPI gene was applied to two important viral serotypes: A and O. Several sequence regions were identified to allow the reconstruction of phylogenetic trees equivalent to those derived from the whole VPI gene. The optimal informative regions for sequence windows of 150 to 250 nt were predicted between positions 250 and 550 of the gene. The sequences spanning the 250 nt of the 3' end (positions 400 to 650), extensively used for FMDV phylogenetic analyses, showed a lower informative content. In spite of this, the use of sequences from this region allowed the derivation of phylogenetic trees for type A and type O FMDVs which showed topologies similar to those previously reported for the whole VP1 gene. When the sequences determined for viruses isolated in Argentina, between 1990 and 1993, were included in these analyses, the results obtained revealed features of the circulation of type A and type O viruses in the field, in the months that preceded the eradication of the disease in this country. Type A viruses were closely related to an Argentinean vaccine strain, and defined an independent cluster within this serotype. Among the type O viruses analysed, two groups were distinguished; one was closely related to the South American vaccine strains, while the other was grouped with viruses of the O3 subtype. In addition, a detailed phylogeny for type A FMDV is presented.
Assuntos
Aphthovirus/genética , Capsídeo/genética , Filogenia , Proteínas Virais/genética , Sequência de Aminoácidos , Animais , Aphthovirus/classificação , Sequência de Bases , Capsídeo/química , Proteínas do Capsídeo , DNA Complementar/química , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , RNA Viral/química , RNA Viral/genética , Sorotipagem , Proteínas Virais/análiseRESUMO
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
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.(AU)