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
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| 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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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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WILLETT, B.; HOLSIE, M.J. Feline leukaemia virus: Half a century since its discovery. The Veterinary Journal , v. 195, n. 1, p. 16-23, 2012. WOOLLEY, A.; LANDON, J. Journal of Immunological Methods, v. 178, p. 253-265, 1995. WU, F.; YUAN, X.Y.; LI, J.; CHEN, Y.H. The co-administration of CpG-ODN influenced protective activity of influenza M2e vaccine. Vaccine, v. 27, n. 32, p. 4320-4324, 2009. ZHANG, Y.; PALMER, G. H.; ABBOTT, J. R.; HOWARD, C. J.; HOPE, J. C.; BROWN, W. C. CpG ODN 2006 and IL-12 are comparable for priming Th1 lymphocyte and IgG responses in cattle immunized with a rickettsial outer membrane protein in alum. Vaccine, v. 21, n. 23, p. 3307-3318, 2003. ZHEN, Y.H.;JIN, L.J.; LI, X.Y.; GUO, J.; LI, Z.; ZHANG, B.J. FANG, R.; XU, Y.P. Efficacy of specific egg yolk immunoglobulin (IgY) to bovine mastitis caused by Staphylococcus aureus. 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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 |
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