Avaliação da antigenicidade de proteínas recombinantes de L. (V.) braziliensis

Detalhes bibliográficos
Autor(a) principal: Alves, José Vitor Ferreira
Data de Publicação: 2014
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFG
dARK ID: ark:/38995/0013000007d25
Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/4201
Resumo: Leishmaniasis is a group of diseases with distinct clinical, histopathological and immunological characteristics, caused by parasites belonging to the Leishmania genus. The serological tests used until the moment have several limitations. There are a great interest to identify immunogenic proteins of Leishmania to be tested as potential antigens for the development of techniques for the diagnosis of American cutaneous leishmaniasis (ACL). The aim of this study was to produce and purify the recombinant proteins "Leishmania activated C kinase" (rLACK); "Thiol Specific Antioxidant" (TSA), "Leishmania elongation initiation factor" (LeIF) and "Leishmania braziliensis stress inducible protein 1" (LbSTI) to evaluate the antigenicity. The recombinant proteins were produced by recombinant DNA techniques as described by Salay et al. (2007). To perform the ELISA, L.(V.) braziliensis (MHOM/BR/1975/M2903) and L.(L.) amazonensis (IFLA/BR/67/PH8) species were cultivated and the antigenic extracts and recombinant proteins were used as antigens. Sixty serum samples from patients with ATL assisted at Anuar Auad hospital, Goiânia, Goiás, were assayed, and from them, 45 were from patients with localized cutaneous leishmaniasis (LCL) and 15 were from mucosal leishmaniasis (ML). To analyze the specificity of the response of total extract and the recombinant proteins, sera from patients with other pathologies were tested. The total extract of L. (L.) amazonensis, rLACK, rTSA, rLbSTI and rLeIF showed a sensitivity of 85%, 75%, 70%, 76.7% and 56.1% respectively. The specificity of total extract of L. (L.) amazonensis, rLACK, rTSA, rLbSTI, rLeIF were 72.5%, 80%, 60%, 30% and 62.5% respectively. Thus, these results showed that recombinants proteins and total extract of L. (L.) amazonensis were antigenic and the total extract of L. (L.) amazonensis and rLACK were the antigen that had the best sensibility and specificity.
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spelling Dorta, Miriam Leandrohttp://lattes.cnpq.br/3933395097851681Dorta, Miriam LeandroMoraes, Sandra do LagoDias, Fátima Ribeirohttp://lattes.cnpq.br/2098038721216478Alves, José Vitor Ferreira2015-02-27T17:20:59Z2014-04-30ALVES, J. V. F. Avaliação da antigenicidade de proteínas recombinantes de L. (V.) braziliensis. 2014. 92 f. Dissertação (Mestrado em Medicina Tropical e Saúde Pública) - Universidade Federal de Goiás, Goiânia, 2014.http://repositorio.bc.ufg.br/tede/handle/tede/4201ark:/38995/0013000007d25Leishmaniasis is a group of diseases with distinct clinical, histopathological and immunological characteristics, caused by parasites belonging to the Leishmania genus. The serological tests used until the moment have several limitations. There are a great interest to identify immunogenic proteins of Leishmania to be tested as potential antigens for the development of techniques for the diagnosis of American cutaneous leishmaniasis (ACL). The aim of this study was to produce and purify the recombinant proteins "Leishmania activated C kinase" (rLACK); "Thiol Specific Antioxidant" (TSA), "Leishmania elongation initiation factor" (LeIF) and "Leishmania braziliensis stress inducible protein 1" (LbSTI) to evaluate the antigenicity. The recombinant proteins were produced by recombinant DNA techniques as described by Salay et al. (2007). To perform the ELISA, L.(V.) braziliensis (MHOM/BR/1975/M2903) and L.(L.) amazonensis (IFLA/BR/67/PH8) species were cultivated and the antigenic extracts and recombinant proteins were used as antigens. Sixty serum samples from patients with ATL assisted at Anuar Auad hospital, Goiânia, Goiás, were assayed, and from them, 45 were from patients with localized cutaneous leishmaniasis (LCL) and 15 were from mucosal leishmaniasis (ML). To analyze the specificity of the response of total extract and the recombinant proteins, sera from patients with other pathologies were tested. The total extract of L. (L.) amazonensis, rLACK, rTSA, rLbSTI and rLeIF showed a sensitivity of 85%, 75%, 70%, 76.7% and 56.1% respectively. The specificity of total extract of L. (L.) amazonensis, rLACK, rTSA, rLbSTI, rLeIF were 72.5%, 80%, 60%, 30% and 62.5% respectively. Thus, these results showed that recombinants proteins and total extract of L. (L.) amazonensis were antigenic and the total extract of L. (L.) amazonensis and rLACK were the antigen that had the best sensibility and specificity.As leishmanioses compreendem um grupo de doenças que apresentam características clínicas, histopatológicas e imunológicas distintas, é causada por parasitos intracelulares que pertencem ao gênero Leishmania. Os testes sorológicos utilizados até o presente, para o diagnóstico, apresentam várias limitações e por isto é de grande importância identificar proteínas imunogênicas da Leishmania, para que sejam testadas como potenciais antígenos para o desenvolvimento de técnicas para o diagnóstico da leishmaniose tegumentar americana (LTA). O objetivo deste estudo foi produzir e purificar as proteínas recombinantes “Leishmania activated C kinase” (rLACK); “Thiol Specific Antioxidant” (TSA), “Leishmania elongation initiation factor” (LeIF) e “Leishmania braziliensis stress inducible protein 1” (LbSTI) e avaliar a sua antigenicidade. As proteínas recombinantes foram produzidas pela técnica de DNA recombinante segundo descrito por Salay et al. (2007). Para a realização do ELISA foram cultivadas as espécies L. (V.) braziliensis (MHOM/BR/1975/M2903) e L. (L.) amazonensis (IFLA/BR/67/PH8) e seus extratos antigênicos e as proteínas recombinantes produzidas foram utilizados como antígenos. Foram utilizadas 60 amostras de pacientes com LTA, atendidos no hospital Anuar Auad, Goiânia, Goiás, sendo que destas, 45 eram de pacientes com leishmaniose cutânea localizada (LCL) e 15 eram de leishmaniose mucosa (LM). Para a análise da especificidade foram utilizados o soro de pacientes com outras patologias. O extrato total de L. (L.) amazonensis, rLACK, rTSA, rLbSTI, rLeIF apresentarem sensibilidade de 85%, 75%, 70%, 76,7%, e 56,1% respectivamente. O extrato total de L. (L.) amazonensis, rLACK, rTSA, rLbSTI, rLeIF obtiveram especificidade de 72,5%, 80%, 60%, 30% e 62,5%, respectivamente. Os resultados do presente trabalho demonstraram que as proteínas recombinantes e o extrato total de L. (L.) amazonensis foram antigênicas e o extrato total de L. (L.) amazonensis e a rLACK foram os antígenos que tiveram as melhores sensibilidades e especificidades.Submitted by Erika Demachki (erikademachki@gmail.com) on 2015-02-27T17:20:49Z No. of bitstreams: 2 Dissertação - José Vitor Ferreira Alves - 2014.pdf: 3685107 bytes, checksum: fe9f9fed0534abe30c80c9eb985428e4 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5)Approved for entry into archive by Erika Demachki (erikademachki@gmail.com) on 2015-02-27T17:20:59Z (GMT) No. of bitstreams: 2 Dissertação - José Vitor Ferreira Alves - 2014.pdf: 3685107 bytes, checksum: fe9f9fed0534abe30c80c9eb985428e4 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5)Made available in DSpace on 2015-02-27T17:20:59Z (GMT). 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dc.title.eng.fl_str_mv Avaliação da antigenicidade de proteínas recombinantes de L. (V.) braziliensis
dc.title.alternative.eng.fl_str_mv Evaluation of antigenicity of recombinant proteins of L. (V.) braziliensis
title Avaliação da antigenicidade de proteínas recombinantes de L. (V.) braziliensis
spellingShingle Avaliação da antigenicidade de proteínas recombinantes de L. (V.) braziliensis
Alves, José Vitor Ferreira
ELISA
Leishmania
Proteínas recombinantes
LTA
Imunodiagnóstico
Immunodiagnostic
Recombinant proteins
ATL
CIENCIAS BIOLOGICAS::IMUNOLOGIA
title_short Avaliação da antigenicidade de proteínas recombinantes de L. (V.) braziliensis
title_full Avaliação da antigenicidade de proteínas recombinantes de L. (V.) braziliensis
title_fullStr Avaliação da antigenicidade de proteínas recombinantes de L. (V.) braziliensis
title_full_unstemmed Avaliação da antigenicidade de proteínas recombinantes de L. (V.) braziliensis
title_sort Avaliação da antigenicidade de proteínas recombinantes de L. (V.) braziliensis
author Alves, José Vitor Ferreira
author_facet Alves, José Vitor Ferreira
author_role author
dc.contributor.advisor1.fl_str_mv Dorta, Miriam Leandro
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/3933395097851681
dc.contributor.referee1.fl_str_mv Dorta, Miriam Leandro
dc.contributor.referee2.fl_str_mv Moraes, Sandra do Lago
dc.contributor.referee3.fl_str_mv Dias, Fátima Ribeiro
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/2098038721216478
dc.contributor.author.fl_str_mv Alves, José Vitor Ferreira
contributor_str_mv Dorta, Miriam Leandro
Dorta, Miriam Leandro
Moraes, Sandra do Lago
Dias, Fátima Ribeiro
dc.subject.por.fl_str_mv ELISA
Leishmania
Proteínas recombinantes
LTA
Imunodiagnóstico
topic ELISA
Leishmania
Proteínas recombinantes
LTA
Imunodiagnóstico
Immunodiagnostic
Recombinant proteins
ATL
CIENCIAS BIOLOGICAS::IMUNOLOGIA
dc.subject.eng.fl_str_mv Immunodiagnostic
Recombinant proteins
ATL
dc.subject.cnpq.fl_str_mv CIENCIAS BIOLOGICAS::IMUNOLOGIA
description Leishmaniasis is a group of diseases with distinct clinical, histopathological and immunological characteristics, caused by parasites belonging to the Leishmania genus. The serological tests used until the moment have several limitations. There are a great interest to identify immunogenic proteins of Leishmania to be tested as potential antigens for the development of techniques for the diagnosis of American cutaneous leishmaniasis (ACL). The aim of this study was to produce and purify the recombinant proteins "Leishmania activated C kinase" (rLACK); "Thiol Specific Antioxidant" (TSA), "Leishmania elongation initiation factor" (LeIF) and "Leishmania braziliensis stress inducible protein 1" (LbSTI) to evaluate the antigenicity. The recombinant proteins were produced by recombinant DNA techniques as described by Salay et al. (2007). To perform the ELISA, L.(V.) braziliensis (MHOM/BR/1975/M2903) and L.(L.) amazonensis (IFLA/BR/67/PH8) species were cultivated and the antigenic extracts and recombinant proteins were used as antigens. Sixty serum samples from patients with ATL assisted at Anuar Auad hospital, Goiânia, Goiás, were assayed, and from them, 45 were from patients with localized cutaneous leishmaniasis (LCL) and 15 were from mucosal leishmaniasis (ML). To analyze the specificity of the response of total extract and the recombinant proteins, sera from patients with other pathologies were tested. The total extract of L. (L.) amazonensis, rLACK, rTSA, rLbSTI and rLeIF showed a sensitivity of 85%, 75%, 70%, 76.7% and 56.1% respectively. The specificity of total extract of L. (L.) amazonensis, rLACK, rTSA, rLbSTI, rLeIF were 72.5%, 80%, 60%, 30% and 62.5% respectively. Thus, these results showed that recombinants proteins and total extract of L. (L.) amazonensis were antigenic and the total extract of L. (L.) amazonensis and rLACK were the antigen that had the best sensibility and specificity.
publishDate 2014
dc.date.issued.fl_str_mv 2014-04-30
dc.date.accessioned.fl_str_mv 2015-02-27T17:20:59Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.citation.fl_str_mv ALVES, J. V. F. Avaliação da antigenicidade de proteínas recombinantes de L. (V.) braziliensis. 2014. 92 f. Dissertação (Mestrado em Medicina Tropical e Saúde Pública) - Universidade Federal de Goiás, Goiânia, 2014.
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identifier_str_mv ALVES, J. V. F. Avaliação da antigenicidade de proteínas recombinantes de L. (V.) braziliensis. 2014. 92 f. Dissertação (Mestrado em Medicina Tropical e Saúde Pública) - Universidade Federal de Goiás, Goiânia, 2014.
ark:/38995/0013000007d25
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dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Instituto de Patologia Tropical e Saúde Pública - IPTSP (RG)
publisher.none.fl_str_mv Universidade Federal de Goiás
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bitstream.checksumAlgorithm.fl_str_mv MD5
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repository.name.fl_str_mv Repositório Institucional da UFG - Universidade Federal de Goiás (UFG)
repository.mail.fl_str_mv tasesdissertacoes.bc@ufg.br
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