Imunoglobulina Y anti-metacestódeo de Taenia crassiceps: ferramenta promissora no imunodiagnóstico da neurocisticercose humana
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFU |
Texto Completo: | https://repositorio.ufu.br/handle/123456789/31416 http://doi.org/10.14393/ufu.te.2020.59 |
Resumo: | Neurocysticercosis (NCC) is a neglected tropical disease and its diagnosis remains a challenge due to nonspecific manifestations. Neuroimaging techniques are used in NCC diagnosis, however, due to the high cost of these methods and the advantages presented in the use of immunological tests, such as easiness to perform and satisfactory results, immunoassays are commonly used to detect antibodies against Taenia sp. antigens. The aim of this study was to produce, fractionate, isolate and characterize IgY antibodies from egg yolk of hens immunized with T. crassiceps metacestodes. Enzyme-linked immunosorbent assay (ELISA) indirect type, avidity ELISA, immunoblotting and indirect immunofluorescence tests were performed for characterization of IgY antibodies. ELISA index increased considerably from the week four, with growth of the production, and high avidity index (AI) were observed; IgY recognized immunodominant polypeptides bands with 24-215 kDa and immunofluorescence assays confirmed the potential of anti-T. crassiceps IgY antibodies to recognize T. crassiceps and T. solium metacestodes tissue. Diagnostic performance was verified by ELISA for immune complexes detection testing 90 serum samples divided into three groups: group 1 (n=30), individuals with NCC definitive diagnosis; group 2 (n = 30), individuals with other parasitic infections; and group 3 (n = 30) apparently healthy individuals. Values of sensitivity (Se), specificity (Sp), positive and negative likelihood ratios (LR+/LR-) and area under the curve (AUC) were calculated. Diagnostic parameters presented the following results: Se 83.3%; Sp 96.7%; LR+ = 25.0; LR− = 0.17 and AUC 0.966. For selection of peptides with binding affinity to anti-Tc and anti-dTc IgY antibodies by phage-display, a library of random peptides expressed in filamentous bacteriophages was used. Three main phage clones were identified, being C11 considered with the best diagnostic values for IgG detection in immunodiagnostic (C11 – Se 80%, Es 83.3%). Results of this study demonstrate that anti-T. crassiceps IgY antibodies present potential applicability and can be used as a promising tool in human NCC immunodiagnosis. |
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Imunoglobulina Y anti-metacestódeo de Taenia crassiceps: ferramenta promissora no imunodiagnóstico da neurocisticercose humanaAnti-Taenia crassiceps metacestode immunoglobulin Y: promising tool in the immunodiagnosis of human neurocysticercosisNeurocisticercoseTaenia crassicepsIgY de gema de ovoImunodiagnósticoNeurocysticercosisegg yolk IgYImmunodiagnosisCNPQ::CIENCIAS BIOLOGICASTestes imunológicosImunoglobulinasNeurocysticercosis (NCC) is a neglected tropical disease and its diagnosis remains a challenge due to nonspecific manifestations. Neuroimaging techniques are used in NCC diagnosis, however, due to the high cost of these methods and the advantages presented in the use of immunological tests, such as easiness to perform and satisfactory results, immunoassays are commonly used to detect antibodies against Taenia sp. antigens. The aim of this study was to produce, fractionate, isolate and characterize IgY antibodies from egg yolk of hens immunized with T. crassiceps metacestodes. Enzyme-linked immunosorbent assay (ELISA) indirect type, avidity ELISA, immunoblotting and indirect immunofluorescence tests were performed for characterization of IgY antibodies. ELISA index increased considerably from the week four, with growth of the production, and high avidity index (AI) were observed; IgY recognized immunodominant polypeptides bands with 24-215 kDa and immunofluorescence assays confirmed the potential of anti-T. crassiceps IgY antibodies to recognize T. crassiceps and T. solium metacestodes tissue. Diagnostic performance was verified by ELISA for immune complexes detection testing 90 serum samples divided into three groups: group 1 (n=30), individuals with NCC definitive diagnosis; group 2 (n = 30), individuals with other parasitic infections; and group 3 (n = 30) apparently healthy individuals. Values of sensitivity (Se), specificity (Sp), positive and negative likelihood ratios (LR+/LR-) and area under the curve (AUC) were calculated. Diagnostic parameters presented the following results: Se 83.3%; Sp 96.7%; LR+ = 25.0; LR− = 0.17 and AUC 0.966. For selection of peptides with binding affinity to anti-Tc and anti-dTc IgY antibodies by phage-display, a library of random peptides expressed in filamentous bacteriophages was used. Three main phage clones were identified, being C11 considered with the best diagnostic values for IgG detection in immunodiagnostic (C11 – Se 80%, Es 83.3%). Results of this study demonstrate that anti-T. crassiceps IgY antibodies present potential applicability and can be used as a promising tool in human NCC immunodiagnosis.FAPEMIG - Fundação de Amparo a Pesquisa do Estado de Minas GeraisTese (Doutorado)A neurocisticercose (NCC) é uma doença tropical negligenciada e seu diagnóstico continua um desafio em vista das manifestações inespecíficas. Técnicas de neuroimagem são utilizadas no diagnóstico da NCC, no entanto, devido ao alto custo desses métodos e as vantagens apresentadas no uso de testes imunológicos, como facilidade de execução e resultados satisfatórios, os imunoensaios são comumente utilizados para detectar anticorpos contra antígenos de Taenia sp. O objetivo deste estudo foi produzir, fracionar, isolar e caracterizar anticorpos IgY provenientes de gemas de ovos de galinhas inoculadas com extrato salino total de formas metacestódeas de T. crassiceps. Ensaios de imunoabsorção enzimática (ELISA) do tipo indireto, ELISA de avidez, immunoblotting e imunofluorescência indireta foram realizados para caracterização dos anticorpos IgY. O índice ELISA aumentou consideravelmente a partir da semana quatro, com crescimento de produção, e observou-se alto índice de avidez (IA); IgY reconheceu bandas polipeptídicas imunodominantes com 24-215 kDa e ensaios de imunofluorescência confirmaram o potencial de anticorpos IgY anti-T. crassiceps para reconhecer tecidos de metacestódeos de T. crassiceps e T. solium. A performance diagnóstica foi verificada por ELISA para a detecção de imunocomplexos, testando 90 amostras de soro divididas em três grupos: grupo 1 (n = 30), indivíduos com diagnóstico definitivo de NCC; grupo 2 (n = 30), indivíduos com outras infecções parasitárias; e grupo 3 (n = 30) indivíduos aparentemente saudáveis. Valores de sensibilidade (Se), especificidade (Sp), razão de probabilidade positiva e negativa (RP+/RP-) e área sob a curva (ASC) foram calculados. Os parâmetros diagnósticos apresentaram os seguintes resultados: Se 83,3%; Es 96,7%; RP + = 25,0; RP− = 0,17 e ASC 0,966. Para seleção de peptídeos com afinidade de ligação aos anticorpos IgY anti-Tc e IgY anti-dTc por phage-display, uma biblioteca de peptídeos randômicos expressados em bacteriófagos filamentosos foi utilizada. Foram identificados três clones de fagos principais, sendo C11 considerado com os melhores valores diagnósticos para detecção de IgG no imunodiagnóstico da NCC humana (C11 - Se 80%, Es 83,3%). Os resultados deste estudo demonstram que anticorpos IgY anti-T. crassiceps apresentam potencial aplicabilidade e podem ser utilizados como uma ferramenta promissora no imunodiagnóstico da NCC humana.2022-02-14Universidade Federal de UberlândiaBrasilPrograma de Pós-graduação em Imunologia e Parasitologia AplicadasCruz, Julia Maria Costahttp://lattes.cnpq.br/2275947687770740Oliveira, Heliana Batista dehttp://lattes.cnpq.br/5388175315143145Faria, Lucas Silva dehttp://lattes.cnpq.br/2802312904104116Miranda, Juliana Silvahttp://lattes.cnpq.br/1892012504907693Ribeiro, Vanessa da Silvahttp://lattes.cnpq.br/4600041017794225Silva, Gabriela Borges da2021-03-11T17:34:12Z2021-03-11T17:34:12Z2020-02-14info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisapplication/pdfSILVA, Gabriela Borges da. Imunoglobulina Y anti-metacestódeo de Taenia crassiceps: ferramenta promissora no imunodiagnóstico da neurocisticercose humana. 202. 106 f. Tese (Doutorado em Imunologia e Parasitologia Aplicadas) - Universidade Federal de Uberlândia, Uberlândia, 2020. DOI http://doi.org/10.14393/ufu.te.2020.59https://repositorio.ufu.br/handle/123456789/31416http://doi.org/10.14393/ufu.te.2020.59porhttp://creativecommons.org/licenses/by-nc-nd/3.0/us/info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFUinstname:Universidade Federal de Uberlândia (UFU)instacron:UFU2022-10-18T13:32:44Zoai:repositorio.ufu.br:123456789/31416Repositório InstitucionalONGhttp://repositorio.ufu.br/oai/requestdiinf@dirbi.ufu.bropendoar:2022-10-18T13:32:44Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)false |
dc.title.none.fl_str_mv |
Imunoglobulina Y anti-metacestódeo de Taenia crassiceps: ferramenta promissora no imunodiagnóstico da neurocisticercose humana Anti-Taenia crassiceps metacestode immunoglobulin Y: promising tool in the immunodiagnosis of human neurocysticercosis |
title |
Imunoglobulina Y anti-metacestódeo de Taenia crassiceps: ferramenta promissora no imunodiagnóstico da neurocisticercose humana |
spellingShingle |
Imunoglobulina Y anti-metacestódeo de Taenia crassiceps: ferramenta promissora no imunodiagnóstico da neurocisticercose humana Silva, Gabriela Borges da Neurocisticercose Taenia crassiceps IgY de gema de ovo Imunodiagnóstico Neurocysticercosis egg yolk IgY Immunodiagnosis CNPQ::CIENCIAS BIOLOGICAS Testes imunológicos Imunoglobulinas |
title_short |
Imunoglobulina Y anti-metacestódeo de Taenia crassiceps: ferramenta promissora no imunodiagnóstico da neurocisticercose humana |
title_full |
Imunoglobulina Y anti-metacestódeo de Taenia crassiceps: ferramenta promissora no imunodiagnóstico da neurocisticercose humana |
title_fullStr |
Imunoglobulina Y anti-metacestódeo de Taenia crassiceps: ferramenta promissora no imunodiagnóstico da neurocisticercose humana |
title_full_unstemmed |
Imunoglobulina Y anti-metacestódeo de Taenia crassiceps: ferramenta promissora no imunodiagnóstico da neurocisticercose humana |
title_sort |
Imunoglobulina Y anti-metacestódeo de Taenia crassiceps: ferramenta promissora no imunodiagnóstico da neurocisticercose humana |
author |
Silva, Gabriela Borges da |
author_facet |
Silva, Gabriela Borges da |
author_role |
author |
dc.contributor.none.fl_str_mv |
Cruz, Julia Maria Costa http://lattes.cnpq.br/2275947687770740 Oliveira, Heliana Batista de http://lattes.cnpq.br/5388175315143145 Faria, Lucas Silva de http://lattes.cnpq.br/2802312904104116 Miranda, Juliana Silva http://lattes.cnpq.br/1892012504907693 Ribeiro, Vanessa da Silva http://lattes.cnpq.br/4600041017794225 |
dc.contributor.author.fl_str_mv |
Silva, Gabriela Borges da |
dc.subject.por.fl_str_mv |
Neurocisticercose Taenia crassiceps IgY de gema de ovo Imunodiagnóstico Neurocysticercosis egg yolk IgY Immunodiagnosis CNPQ::CIENCIAS BIOLOGICAS Testes imunológicos Imunoglobulinas |
topic |
Neurocisticercose Taenia crassiceps IgY de gema de ovo Imunodiagnóstico Neurocysticercosis egg yolk IgY Immunodiagnosis CNPQ::CIENCIAS BIOLOGICAS Testes imunológicos Imunoglobulinas |
description |
Neurocysticercosis (NCC) is a neglected tropical disease and its diagnosis remains a challenge due to nonspecific manifestations. Neuroimaging techniques are used in NCC diagnosis, however, due to the high cost of these methods and the advantages presented in the use of immunological tests, such as easiness to perform and satisfactory results, immunoassays are commonly used to detect antibodies against Taenia sp. antigens. The aim of this study was to produce, fractionate, isolate and characterize IgY antibodies from egg yolk of hens immunized with T. crassiceps metacestodes. Enzyme-linked immunosorbent assay (ELISA) indirect type, avidity ELISA, immunoblotting and indirect immunofluorescence tests were performed for characterization of IgY antibodies. ELISA index increased considerably from the week four, with growth of the production, and high avidity index (AI) were observed; IgY recognized immunodominant polypeptides bands with 24-215 kDa and immunofluorescence assays confirmed the potential of anti-T. crassiceps IgY antibodies to recognize T. crassiceps and T. solium metacestodes tissue. Diagnostic performance was verified by ELISA for immune complexes detection testing 90 serum samples divided into three groups: group 1 (n=30), individuals with NCC definitive diagnosis; group 2 (n = 30), individuals with other parasitic infections; and group 3 (n = 30) apparently healthy individuals. Values of sensitivity (Se), specificity (Sp), positive and negative likelihood ratios (LR+/LR-) and area under the curve (AUC) were calculated. Diagnostic parameters presented the following results: Se 83.3%; Sp 96.7%; LR+ = 25.0; LR− = 0.17 and AUC 0.966. For selection of peptides with binding affinity to anti-Tc and anti-dTc IgY antibodies by phage-display, a library of random peptides expressed in filamentous bacteriophages was used. Three main phage clones were identified, being C11 considered with the best diagnostic values for IgG detection in immunodiagnostic (C11 – Se 80%, Es 83.3%). Results of this study demonstrate that anti-T. crassiceps IgY antibodies present potential applicability and can be used as a promising tool in human NCC immunodiagnosis. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-02-14 2021-03-11T17:34:12Z 2021-03-11T17:34:12Z |
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.uri.fl_str_mv |
SILVA, Gabriela Borges da. Imunoglobulina Y anti-metacestódeo de Taenia crassiceps: ferramenta promissora no imunodiagnóstico da neurocisticercose humana. 202. 106 f. Tese (Doutorado em Imunologia e Parasitologia Aplicadas) - Universidade Federal de Uberlândia, Uberlândia, 2020. DOI http://doi.org/10.14393/ufu.te.2020.59 https://repositorio.ufu.br/handle/123456789/31416 http://doi.org/10.14393/ufu.te.2020.59 |
identifier_str_mv |
SILVA, Gabriela Borges da. Imunoglobulina Y anti-metacestódeo de Taenia crassiceps: ferramenta promissora no imunodiagnóstico da neurocisticercose humana. 202. 106 f. Tese (Doutorado em Imunologia e Parasitologia Aplicadas) - Universidade Federal de Uberlândia, Uberlândia, 2020. DOI http://doi.org/10.14393/ufu.te.2020.59 |
url |
https://repositorio.ufu.br/handle/123456789/31416 http://doi.org/10.14393/ufu.te.2020.59 |
dc.language.iso.fl_str_mv |
por |
language |
por |
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http://creativecommons.org/licenses/by-nc-nd/3.0/us/ info:eu-repo/semantics/openAccess |
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http://creativecommons.org/licenses/by-nc-nd/3.0/us/ |
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openAccess |
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application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Uberlândia Brasil Programa de Pós-graduação em Imunologia e Parasitologia Aplicadas |
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Universidade Federal de Uberlândia Brasil Programa de Pós-graduação em Imunologia e Parasitologia Aplicadas |
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reponame:Repositório Institucional da UFU instname:Universidade Federal de Uberlândia (UFU) instacron:UFU |
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Universidade Federal de Uberlândia (UFU) |
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UFU |
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Repositório Institucional da UFU |
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Repositório Institucional da UFU |
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Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU) |
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diinf@dirbi.ufu.br |
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