Caracterização genética e antigênica de isolados brasileiros do vírus da diarréia viral bovina

Detalhes bibliográficos
Autor(a) principal: Bianchi, Eloisa
Data de Publicação: 2011
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
dARK ID: ark:/26339/00130000017nj
Texto Completo: http://repositorio.ufsm.br/handle/1/10089
Resumo: Bovine viral diarrhea virus (BVDV) isolates display a high genetic and antigenic variability that can compromise diagnosis and vaccine formulation. This study aimed to characterize at genetic and antigenic levels Brazilian BVDV isolates for potential use in diagnosis and vaccine production, and to characterize field BVDV isolates from Rio Grande do Sul (RS) (2000-2010). Chapter 1 describes the genetic and antigenic characterization of seven cytopathic Brazilian BVDV isolates, to select the most appropriate for use in diagnostic and vaccines. Sequencing and phylogeny identified four isolates of BVDV-1 (57.1%) and three isolates of BVDV-2 genotype (42.8%). The antigenic characterization of these isolates by virus neutralization assay (VN) with homologous and heterologous antisera revealed differences among the viruses of the same genotype, but especially among viruses of different genotypes. Despite the antigenic differences, the neutralizing activity of antisera IBSP-4 and VS26/2 was acceptable against isolates of the same genotype and also against viruses of different genotype. These results qualify these isolates for potential use in vaccine formulations. In parallel, 446 field serum samples were tested against the seven isolates and reference strains. Neutralizing antibodies anti-BVDV were found in 221 (50.3%) samples, and 198 (89.6%) had neutralizing activity against the both genotypes (Singer, BVDV-1 and VS- 253, BVDV-2), 18 (8.1%) serum samples and five (2.2%) neutralized only the standard BVDV-1 and BVDV-2. These results confirm that the antigenic differences between isolates may result in false-negative results, yet might not justify the performance of VN using both BVDV genotypes. Chapter 2 presents a genotypic and antigenic profile of 20 BVDV isolates obtained in RS state, between 2000 and 2010. The isolates were isolated from a variety of clinico-pathological syndromes. Most samples (19 or 95%) belong to biotype non-cytopathic (NCP) and one isolate (5%) had a mixture of viruses NCP and cytopathic (CP). Sequencing and phylogenetic analyses identified nine isolates of BVDV-2 (45%), eight BVDV-1 (40%) and three atypical BVDV isolates. Analysis of reactivity with a panel of 20 monoclonal antibodies (mAbs) revealed a marked variability in the major envelope glycoprotein gp53/E2 among viruses of the same genotype, but especially between different virus genotypes. Virus neutralization assays (VN) with antisera of BVDV-1 and BVDV-2 reference strains against the isolates revealed variable levels of cross-reactivity between viruses of the same genotype, and lack or very low reactivity between viruses of different genotypes. These results indicate the presence of both genotypes of BVDV in the cattle population of RS and confirm the remarkable antigenic diversity of these isolates.
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spelling Caracterização genética e antigênica de isolados brasileiros do vírus da diarréia viral bovinaGenetic and antigenic characterization of brazilian isolates of bovine viral diarrhea virus isolatesBVDV-1BVDV-2PestivirusAnálise filogenéticaAnálise antigênicaBVDV-1BVDV-2PestivirusPhylogenetic analysisAntigenic analysisCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIABovine viral diarrhea virus (BVDV) isolates display a high genetic and antigenic variability that can compromise diagnosis and vaccine formulation. This study aimed to characterize at genetic and antigenic levels Brazilian BVDV isolates for potential use in diagnosis and vaccine production, and to characterize field BVDV isolates from Rio Grande do Sul (RS) (2000-2010). Chapter 1 describes the genetic and antigenic characterization of seven cytopathic Brazilian BVDV isolates, to select the most appropriate for use in diagnostic and vaccines. Sequencing and phylogeny identified four isolates of BVDV-1 (57.1%) and three isolates of BVDV-2 genotype (42.8%). The antigenic characterization of these isolates by virus neutralization assay (VN) with homologous and heterologous antisera revealed differences among the viruses of the same genotype, but especially among viruses of different genotypes. Despite the antigenic differences, the neutralizing activity of antisera IBSP-4 and VS26/2 was acceptable against isolates of the same genotype and also against viruses of different genotype. These results qualify these isolates for potential use in vaccine formulations. In parallel, 446 field serum samples were tested against the seven isolates and reference strains. Neutralizing antibodies anti-BVDV were found in 221 (50.3%) samples, and 198 (89.6%) had neutralizing activity against the both genotypes (Singer, BVDV-1 and VS- 253, BVDV-2), 18 (8.1%) serum samples and five (2.2%) neutralized only the standard BVDV-1 and BVDV-2. These results confirm that the antigenic differences between isolates may result in false-negative results, yet might not justify the performance of VN using both BVDV genotypes. Chapter 2 presents a genotypic and antigenic profile of 20 BVDV isolates obtained in RS state, between 2000 and 2010. The isolates were isolated from a variety of clinico-pathological syndromes. Most samples (19 or 95%) belong to biotype non-cytopathic (NCP) and one isolate (5%) had a mixture of viruses NCP and cytopathic (CP). Sequencing and phylogenetic analyses identified nine isolates of BVDV-2 (45%), eight BVDV-1 (40%) and three atypical BVDV isolates. Analysis of reactivity with a panel of 20 monoclonal antibodies (mAbs) revealed a marked variability in the major envelope glycoprotein gp53/E2 among viruses of the same genotype, but especially between different virus genotypes. Virus neutralization assays (VN) with antisera of BVDV-1 and BVDV-2 reference strains against the isolates revealed variable levels of cross-reactivity between viruses of the same genotype, and lack or very low reactivity between viruses of different genotypes. These results indicate the presence of both genotypes of BVDV in the cattle population of RS and confirm the remarkable antigenic diversity of these isolates.Conselho Nacional de Desenvolvimento Científico e TecnológicoAmostras de campo do vírus da diarréia viral bovina (BVDV) apresentam diferenças genéticas e antigênicas que podem influenciar no diagnóstico e na produção de vacinas. Os objetivos do presente trabalho foram selecionar e caracterizar isolados de BVDV para potencial uso em testes de diagnóstico e vacinas, e caracterizar genética e antigenicamente amostras de BVDV isoladas no Rio Grande do Sul (RS) entre 2000 e 2010. O capítulo 1 relata a caracterização em nível molecular e antigênico de sete isolados citopáticos brasileiros do BVDV. O sequenciamento e análise filogenética identificaram quatro isolados do genótipo BVDV-1 (57,1%) e três isolados do genótipo BVDV-2 (42,8%). A caracterização antigênica dessas amostras por vírus neutralização (VN) com soro homólogo e heterólogo revelou diferenças importantes nos títulos neutralizantes entre os vírus do mesmo genótipo, mas principalmente entre vírus de diferentes genótipos. Apesar dessas diferenças antigênicas, a atividade neutralizante do anti-soro das amostras IBSP-4 e VS26/2 foi aceitável contra amostras do mesmo genótipo e também contra amostras de genótipo diferente. Esses resultados qualificam esses isolados para potencial uso em formulações vacinais. Em paralelo, 446 amostras de soro campo foram testadas por VN frente aos sete isolados e contra cepas padrão de BVDV-1 e 2. Destas, 221 amostras (50,3%) apresentaram anticorpos neutralizantes anti-BVDV, sendo que 198 (89,6%) apresentaram atividade frente às cepas padrões dos dois genótipos (Singer, BVDV-1 e VS-253, BVDV-2). Por outro lado, 18 amostras de soro (8,1%) neutralizaram apenas a cepa de BVDV-1 e 5 (2,2%) a cepa BVDV-2. Apesar das diferenças antigênicas entre os genótipos, esses resultados não indicam a necessidade de testar as amostras de soro para ambos os genótipos, uma vez que o número de amostras de soro que não é detectada por um ou outro genótipo foi pequena. O capítulo 2 apresenta um perfil genotípico e antigênico de 20 amostras do BVDV isoladas no RS, entre 2000 e 2010. As amostras foram isoladas de uma variedade de condições clínicas. A maioria das amostras (19 ou 95%) pertence ao biótipo não-citopático (NCP); e um isolado (5%) apresentou uma mistura de vírus NCP e citopáticos (CP). O sequenciamento e a análise filogenética permitiram identificar nove isolados de BVDV-2 (45%), oito isolados de BVDV-1 (40%) e três isolados BVDV atípicos. Análise de reatividade com um painel de 20 anticorpos monoclonais (AcMs) contra a glicoproteína principal do envelope gp53/E2 revelou uma variabilidade marcante nesta glicoproteína, entre vírus do mesmo genótipo, e sobretudo, entre vírus de genótipos diferentes. Testes de VN com anti-soro de cepas de referência de BVDV-1 e BVDV-2 frente às amostras isoladas revelaram níveis variáveis de reatividade cruzada entre vírus do mesmo genótipo, e reatividade ausente ou muito baixa entre vírus de genótipos diferentes. Esses resultados indicam a presença de ambos os genótipos do BVDV na população do RS, e confirmam a marcante variabilidade antigênica desses isolados.Universidade Federal de Santa MariaBRMedicina VeterináriaUFSMPrograma de Pós-Graduação em Medicina VeterináriaWeiblen, Rudihttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783394D5Flores, Eduardo Furtadohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4785140A1Scherer, Charles Fernando Capinoshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4767498P7Bianchi, Eloisa2017-06-012017-06-012011-02-16info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfBIANCHI, Eloisa. Genetic and antigenic characterization of brazilian isolates of bovine viral diarrhea virus isolates. 2011. 60 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2011.http://repositorio.ufsm.br/handle/1/10089ark:/26339/00130000017njporinfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSM2017-07-25T15:01:32Zoai:repositorio.ufsm.br:1/10089Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2017-07-25T15:01:32Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.none.fl_str_mv Caracterização genética e antigênica de isolados brasileiros do vírus da diarréia viral bovina
Genetic and antigenic characterization of brazilian isolates of bovine viral diarrhea virus isolates
title Caracterização genética e antigênica de isolados brasileiros do vírus da diarréia viral bovina
spellingShingle Caracterização genética e antigênica de isolados brasileiros do vírus da diarréia viral bovina
Bianchi, Eloisa
BVDV-1
BVDV-2
Pestivirus
Análise filogenética
Análise antigênica
BVDV-1
BVDV-2
Pestivirus
Phylogenetic analysis
Antigenic analysis
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Caracterização genética e antigênica de isolados brasileiros do vírus da diarréia viral bovina
title_full Caracterização genética e antigênica de isolados brasileiros do vírus da diarréia viral bovina
title_fullStr Caracterização genética e antigênica de isolados brasileiros do vírus da diarréia viral bovina
title_full_unstemmed Caracterização genética e antigênica de isolados brasileiros do vírus da diarréia viral bovina
title_sort Caracterização genética e antigênica de isolados brasileiros do vírus da diarréia viral bovina
author Bianchi, Eloisa
author_facet Bianchi, Eloisa
author_role author
dc.contributor.none.fl_str_mv Weiblen, Rudi
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783394D5
Flores, Eduardo Furtado
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4785140A1
Scherer, Charles Fernando Capinos
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4767498P7
dc.contributor.author.fl_str_mv Bianchi, Eloisa
dc.subject.por.fl_str_mv BVDV-1
BVDV-2
Pestivirus
Análise filogenética
Análise antigênica
BVDV-1
BVDV-2
Pestivirus
Phylogenetic analysis
Antigenic analysis
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
topic BVDV-1
BVDV-2
Pestivirus
Análise filogenética
Análise antigênica
BVDV-1
BVDV-2
Pestivirus
Phylogenetic analysis
Antigenic analysis
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description Bovine viral diarrhea virus (BVDV) isolates display a high genetic and antigenic variability that can compromise diagnosis and vaccine formulation. This study aimed to characterize at genetic and antigenic levels Brazilian BVDV isolates for potential use in diagnosis and vaccine production, and to characterize field BVDV isolates from Rio Grande do Sul (RS) (2000-2010). Chapter 1 describes the genetic and antigenic characterization of seven cytopathic Brazilian BVDV isolates, to select the most appropriate for use in diagnostic and vaccines. Sequencing and phylogeny identified four isolates of BVDV-1 (57.1%) and three isolates of BVDV-2 genotype (42.8%). The antigenic characterization of these isolates by virus neutralization assay (VN) with homologous and heterologous antisera revealed differences among the viruses of the same genotype, but especially among viruses of different genotypes. Despite the antigenic differences, the neutralizing activity of antisera IBSP-4 and VS26/2 was acceptable against isolates of the same genotype and also against viruses of different genotype. These results qualify these isolates for potential use in vaccine formulations. In parallel, 446 field serum samples were tested against the seven isolates and reference strains. Neutralizing antibodies anti-BVDV were found in 221 (50.3%) samples, and 198 (89.6%) had neutralizing activity against the both genotypes (Singer, BVDV-1 and VS- 253, BVDV-2), 18 (8.1%) serum samples and five (2.2%) neutralized only the standard BVDV-1 and BVDV-2. These results confirm that the antigenic differences between isolates may result in false-negative results, yet might not justify the performance of VN using both BVDV genotypes. Chapter 2 presents a genotypic and antigenic profile of 20 BVDV isolates obtained in RS state, between 2000 and 2010. The isolates were isolated from a variety of clinico-pathological syndromes. Most samples (19 or 95%) belong to biotype non-cytopathic (NCP) and one isolate (5%) had a mixture of viruses NCP and cytopathic (CP). Sequencing and phylogenetic analyses identified nine isolates of BVDV-2 (45%), eight BVDV-1 (40%) and three atypical BVDV isolates. Analysis of reactivity with a panel of 20 monoclonal antibodies (mAbs) revealed a marked variability in the major envelope glycoprotein gp53/E2 among viruses of the same genotype, but especially between different virus genotypes. Virus neutralization assays (VN) with antisera of BVDV-1 and BVDV-2 reference strains against the isolates revealed variable levels of cross-reactivity between viruses of the same genotype, and lack or very low reactivity between viruses of different genotypes. These results indicate the presence of both genotypes of BVDV in the cattle population of RS and confirm the remarkable antigenic diversity of these isolates.
publishDate 2011
dc.date.none.fl_str_mv 2011-02-16
2017-06-01
2017-06-01
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.uri.fl_str_mv BIANCHI, Eloisa. Genetic and antigenic characterization of brazilian isolates of bovine viral diarrhea virus isolates. 2011. 60 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2011.
http://repositorio.ufsm.br/handle/1/10089
dc.identifier.dark.fl_str_mv ark:/26339/00130000017nj
identifier_str_mv BIANCHI, Eloisa. Genetic and antigenic characterization of brazilian isolates of bovine viral diarrhea virus isolates. 2011. 60 f. Dissertação (Mestrado em Medicina Veterinária) - Universidade Federal de Santa Maria, Santa Maria, 2011.
ark:/26339/00130000017nj
url http://repositorio.ufsm.br/handle/1/10089
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Santa Maria
BR
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
publisher.none.fl_str_mv Universidade Federal de Santa Maria
BR
Medicina Veterinária
UFSM
Programa de Pós-Graduação em Medicina Veterinária
dc.source.none.fl_str_mv reponame:Manancial - Repositório Digital da UFSM
instname:Universidade Federal de Santa Maria (UFSM)
instacron:UFSM
instname_str Universidade Federal de Santa Maria (UFSM)
instacron_str UFSM
institution UFSM
reponame_str Manancial - Repositório Digital da UFSM
collection Manancial - Repositório Digital da UFSM
repository.name.fl_str_mv Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)
repository.mail.fl_str_mv atendimento.sib@ufsm.br||tedebc@gmail.com
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