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Previous issue date: 2014-04-30 / 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. / 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.
| Identifer | oai:union.ndltd.org:IBICT/oai:repositorio.bc.ufg.br:tede/4201 |
| Date | 30 April 2014 |
| Creators | Alves, José Vitor Ferreira |
| Contributors | Dorta, Miriam Leandro, Dorta, Miriam Leandro, Moraes, Sandra do Lago, Dias, Fátima Ribeiro |
| Publisher | Universidade Federal de Goiás, Programa de Pós-graduação em Medicina Tropical e Saúde Publica (IPTSP), UFG, Brasil, Instituto de Patologia Tropical e Saúde Pública - IPTSP (RG) |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | Portuguese |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Format | application/pdf |
| Source | reponame:Biblioteca Digital de Teses e Dissertações da UFG, instname:Universidade Federal de Goiás, instacron:UFG |
| Rights | http://creativecommons.org/licenses/by-nc-nd/4.0/, info:eu-repo/semantics/openAccess |
| Relation | 6085308344741430434, 600, 600, 600, -7769011444564556288, 5989919188376747614 |
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