Clonagem e expressão da proteína recombinante p27 (r-p27) do vírus da leucemia felina e produção de IgY aviária contra a r-p27 para utilização em imunodiagnóstico

Detalhes bibliográficos
Autor(a) principal: Almeida, Nadia Rossi de
Data de Publicação: 2013
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFRRJ
Texto Completo: https://rima.ufrrj.br/jspui/handle/20.500.14407/9574
Resumo: O Vírus da Leucemia Felina (FeLV) é um membro da família Retroviridae, gênero Gammaretrovirus, que representa uma importante causa de morbidade e mortalidade entre os gatos domésticos. No Brasil, o acesso ao diagnóstico da Leucemia Viral Felina ainda é restrito devido ao alto custo do teste comercial disponível, que detecta o antígeno viral p27. Para utilização em imunodiagnóstico, a IgY aviária apresenta uma série de vantagens sobre os anticorpos IgG de mamíferos, como o baixo custo da técnica e manutenção dos animais, alto rendimento dos anticorpos purificados e caráter bioético. O objetivo do presente estudo foi o desenvolvimento de uma proteína de nucleocapsídeo viral p27 recombinante, a r-p27, para elicitar anticorpos aviários, visando contribuir com o imunodiagnóstico das infecções pelo FeLV no país. Esta tecnologia poderá possibilitar o acesso ao diagnóstico e a melhor compreensão dos estudos de prevalência da infecção. Para tanto, a região codificante da p27 nativa do FeLV foi amplificada, clonada e expressa em Escherichia coli, e sua reatividade contra os anticorpos para p27 nativa foi verificada com o kit comercial de imunocromatografia. A r-p27 foi inoculada em coelhos e galinhas, para produção de IgG e IgY específicas, respectivamente. Estes anticorpos foram avaliados pela técnica de imunofluorescência indireta (IFI) contra a p27 nativa, em esfregaços de sangue de animais positivos e negativos para o FeLV. Ambos os tipos de imunoglobulinas apresentaram reatividade para a p27 recombinante e nativa do FeLV. A produção de IgY para a r-p27 representou um método de alto rendimento e baixo custo, promissor para o desenvolvimento de imunodiagnóstico da Leucemia Viral Felina
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spelling Almeida, Nadia Rossi deMazur, Carlos610.764.147-53http://lattes.cnpq.br/7059968849728033Danelli, Maria das Graças Miranda758.643.637-15http://lattes.cnpq.br/7414712085904286Hagiwara, Mitika KuribayashiReis, Jenner Karlisson Pimenta dosMcIntosh, DouglasSantos, Débora Regina Lopes dos086.087.827-90http://lattes.cnpq.br/42922268425304902023-12-21T18:41:45Z2023-12-21T18:41:45Z2013-02-20ALMEIDA, Nadia Rossi de. Clonagem e expressão da proteína recombinante p27 (r-p27) do vírus da leucemia felina e produção de IgY aviária contra a r-p27 para utilização em imunodiagnóstico. 2013. 87 f. Tese (Doutorado em Ciências Veterinárias). Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, 2013https://rima.ufrrj.br/jspui/handle/20.500.14407/9574O Vírus da Leucemia Felina (FeLV) é um membro da família Retroviridae, gênero Gammaretrovirus, que representa uma importante causa de morbidade e mortalidade entre os gatos domésticos. No Brasil, o acesso ao diagnóstico da Leucemia Viral Felina ainda é restrito devido ao alto custo do teste comercial disponível, que detecta o antígeno viral p27. Para utilização em imunodiagnóstico, a IgY aviária apresenta uma série de vantagens sobre os anticorpos IgG de mamíferos, como o baixo custo da técnica e manutenção dos animais, alto rendimento dos anticorpos purificados e caráter bioético. O objetivo do presente estudo foi o desenvolvimento de uma proteína de nucleocapsídeo viral p27 recombinante, a r-p27, para elicitar anticorpos aviários, visando contribuir com o imunodiagnóstico das infecções pelo FeLV no país. Esta tecnologia poderá possibilitar o acesso ao diagnóstico e a melhor compreensão dos estudos de prevalência da infecção. Para tanto, a região codificante da p27 nativa do FeLV foi amplificada, clonada e expressa em Escherichia coli, e sua reatividade contra os anticorpos para p27 nativa foi verificada com o kit comercial de imunocromatografia. A r-p27 foi inoculada em coelhos e galinhas, para produção de IgG e IgY específicas, respectivamente. Estes anticorpos foram avaliados pela técnica de imunofluorescência indireta (IFI) contra a p27 nativa, em esfregaços de sangue de animais positivos e negativos para o FeLV. Ambos os tipos de imunoglobulinas apresentaram reatividade para a p27 recombinante e nativa do FeLV. A produção de IgY para a r-p27 representou um método de alto rendimento e baixo custo, promissor para o desenvolvimento de imunodiagnóstico da Leucemia Viral FelinaCoordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESFeline Leukemia Virus (FeLV) is a gammaretrovirus, which represents an important cause of morbidity and mortality among domestic cats. In Brazil, the access to feline leukemia diagnosis is still limited due to the high cost of available commercial test to detect the viral antigen p27. The production of avian IgY instead of mammal IgG has several advantages, like lower costs of the technique, easier animal maintenance, high yields of purified antibodies and better acceptance due to bioethical characteristics. The aim of this study was the development of a recombinant viral p27 protein, the r-p27, to elicit avian antibodies, aiming to enable immunodiagnosis of FeLV infections. This technology can provide access to diagnosis and better understanding of prevalence studies. For this purpose, the coding region of the FeLV p27 protein was cloned and expressed in Escherichia coli and the reactivity against anti-p27 antibody was proven before commercial immunochromatography kit. To obtain IgG and IgY anti-p27, r-p27 was inoculated into rabbits and chickens, respectively, and the antibodies produced were tested by indirect immunofluorescence (IFA). Both immunoglobulins showed reactivity against recombinant and native p27. 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dc.title.por.fl_str_mv Clonagem e expressão da proteína recombinante p27 (r-p27) do vírus da leucemia felina e produção de IgY aviária contra a r-p27 para utilização em imunodiagnóstico
dc.title.alternative.eng.fl_str_mv Cloning and expression of p27 protein (r-p27) of Feline Leukemia Virus and avian IgY production against the r-p27 to immunodiagnostic application
title Clonagem e expressão da proteína recombinante p27 (r-p27) do vírus da leucemia felina e produção de IgY aviária contra a r-p27 para utilização em imunodiagnóstico
spellingShingle Clonagem e expressão da proteína recombinante p27 (r-p27) do vírus da leucemia felina e produção de IgY aviária contra a r-p27 para utilização em imunodiagnóstico
Almeida, Nadia Rossi de
FeLV
imunodiagnóstico
IgY
immunodiagnosis
Medicina Veterinária
title_short Clonagem e expressão da proteína recombinante p27 (r-p27) do vírus da leucemia felina e produção de IgY aviária contra a r-p27 para utilização em imunodiagnóstico
title_full Clonagem e expressão da proteína recombinante p27 (r-p27) do vírus da leucemia felina e produção de IgY aviária contra a r-p27 para utilização em imunodiagnóstico
title_fullStr Clonagem e expressão da proteína recombinante p27 (r-p27) do vírus da leucemia felina e produção de IgY aviária contra a r-p27 para utilização em imunodiagnóstico
title_full_unstemmed Clonagem e expressão da proteína recombinante p27 (r-p27) do vírus da leucemia felina e produção de IgY aviária contra a r-p27 para utilização em imunodiagnóstico
title_sort Clonagem e expressão da proteína recombinante p27 (r-p27) do vírus da leucemia felina e produção de IgY aviária contra a r-p27 para utilização em imunodiagnóstico
author Almeida, Nadia Rossi de
author_facet Almeida, Nadia Rossi de
author_role author
dc.contributor.author.fl_str_mv Almeida, Nadia Rossi de
dc.contributor.advisor1.fl_str_mv Mazur, Carlos
dc.contributor.advisor1ID.fl_str_mv 610.764.147-53
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/7059968849728033
dc.contributor.advisor-co1.fl_str_mv Danelli, Maria das Graças Miranda
dc.contributor.advisor-co1ID.fl_str_mv 758.643.637-15
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/7414712085904286
dc.contributor.referee1.fl_str_mv Hagiwara, Mitika Kuribayashi
dc.contributor.referee2.fl_str_mv Reis, Jenner Karlisson Pimenta dos
dc.contributor.referee3.fl_str_mv McIntosh, Douglas
dc.contributor.referee4.fl_str_mv Santos, Débora Regina Lopes dos
dc.contributor.authorID.fl_str_mv 086.087.827-90
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/4292226842530490
contributor_str_mv Mazur, Carlos
Danelli, Maria das Graças Miranda
Hagiwara, Mitika Kuribayashi
Reis, Jenner Karlisson Pimenta dos
McIntosh, Douglas
Santos, Débora Regina Lopes dos
dc.subject.por.fl_str_mv FeLV
imunodiagnóstico
IgY
topic FeLV
imunodiagnóstico
IgY
immunodiagnosis
Medicina Veterinária
dc.subject.eng.fl_str_mv immunodiagnosis
dc.subject.cnpq.fl_str_mv Medicina Veterinária
description O Vírus da Leucemia Felina (FeLV) é um membro da família Retroviridae, gênero Gammaretrovirus, que representa uma importante causa de morbidade e mortalidade entre os gatos domésticos. No Brasil, o acesso ao diagnóstico da Leucemia Viral Felina ainda é restrito devido ao alto custo do teste comercial disponível, que detecta o antígeno viral p27. Para utilização em imunodiagnóstico, a IgY aviária apresenta uma série de vantagens sobre os anticorpos IgG de mamíferos, como o baixo custo da técnica e manutenção dos animais, alto rendimento dos anticorpos purificados e caráter bioético. O objetivo do presente estudo foi o desenvolvimento de uma proteína de nucleocapsídeo viral p27 recombinante, a r-p27, para elicitar anticorpos aviários, visando contribuir com o imunodiagnóstico das infecções pelo FeLV no país. Esta tecnologia poderá possibilitar o acesso ao diagnóstico e a melhor compreensão dos estudos de prevalência da infecção. Para tanto, a região codificante da p27 nativa do FeLV foi amplificada, clonada e expressa em Escherichia coli, e sua reatividade contra os anticorpos para p27 nativa foi verificada com o kit comercial de imunocromatografia. A r-p27 foi inoculada em coelhos e galinhas, para produção de IgG e IgY específicas, respectivamente. Estes anticorpos foram avaliados pela técnica de imunofluorescência indireta (IFI) contra a p27 nativa, em esfregaços de sangue de animais positivos e negativos para o FeLV. Ambos os tipos de imunoglobulinas apresentaram reatividade para a p27 recombinante e nativa do FeLV. A produção de IgY para a r-p27 representou um método de alto rendimento e baixo custo, promissor para o desenvolvimento de imunodiagnóstico da Leucemia Viral Felina
publishDate 2013
dc.date.issued.fl_str_mv 2013-02-20
dc.date.accessioned.fl_str_mv 2023-12-21T18:41:45Z
dc.date.available.fl_str_mv 2023-12-21T18:41:45Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv ALMEIDA, Nadia Rossi de. Clonagem e expressão da proteína recombinante p27 (r-p27) do vírus da leucemia felina e produção de IgY aviária contra a r-p27 para utilização em imunodiagnóstico. 2013. 87 f. Tese (Doutorado em Ciências Veterinárias). Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, 2013
dc.identifier.uri.fl_str_mv https://rima.ufrrj.br/jspui/handle/20.500.14407/9574
identifier_str_mv ALMEIDA, Nadia Rossi de. Clonagem e expressão da proteína recombinante p27 (r-p27) do vírus da leucemia felina e produção de IgY aviária contra a r-p27 para utilização em imunodiagnóstico. 2013. 87 f. Tese (Doutorado em Ciências Veterinárias). Instituto de Veterinária, Universidade Federal Rural do Rio de Janeiro, Seropédica, 2013
url https://rima.ufrrj.br/jspui/handle/20.500.14407/9574
dc.language.iso.fl_str_mv por
language por
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