Reconhecimento da ligação dos anticorpos anti-HCV com proteínas recombinantes do vírus da hepatite C por meio do teste ELISA
Ano de defesa: | 2016 |
---|---|
Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | |
Tipo de documento: | Tese |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Universidade Federal de São Carlos
Câmpus São Carlos |
Programa de Pós-Graduação: |
Programa de Pós-Graduação em Biotecnologia - PPGBiotec
|
Departamento: |
Não Informado pela instituição
|
País: |
Não Informado pela instituição
|
Palavras-chave em Inglês: | |
Área do conhecimento CNPq: | |
Link de acesso: | https://repositorio.ufscar.br/handle/ufscar/8396 |
Resumo: | Viral hepatitis is a major public health problem worldwide and in Brazil. They are notifiable diseases and according to estimates, billions of people have had contact with the hepatitis and millions are chronic carriers. Infection with hepatitis C virus (VHC) is a major problem worldwide public health due to the high rate of progression to chronicity, the evolutionary potential for cirrhosis and hepatocellular carcinoma, major complications leading to death. In general it can be said that in the last decade there have been major advances in the diagnosis of hepatitis C. In this period there was progressive improvement in sensitivity and specificity of the tests used to detect antibodies against the VHC virus. However, it is necessary that more accurate tests are developed. Thus, considering that the concern for the detection of hepatitis C increases every day, especially in blood banks; the diagnostic methods of this infection are of great clinical relevance and may be used as markers chronicity and indicative of therapeutic efficacy. Therefore, this work proposed to evaluate the connection and recognition of anti-VHC antibody positive and positive genotyped samples in patients with hepatitis C through the standardization of procedures and solutions used in qualitative ELISA. There was the process of awareness of microplates with recombinant chimeric protein, made the analysis of sensitivity, specificity, reproducibility and validity of the method. We obtained from ELISA assays with standardized recombinant proteins a protocol able to have a good performance of the main components of the reaction, and antigens conjugated with good resolution. This study presented ELISA results valid 95.69%, 100% reproducibility, 94.5% sensitivity and specificity 99.3%, higher than the ELISA performed with multiepitopo protein MEHCV who presented with sensitivity (92.86%) and specificity (82.89%). The standard ELISA can be used as a qualitative serological technique aimed at detection of anti-VHC antibodies, as demonstrated with great reactivity in patients infected with VHC. |
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Souza, Laís Cristina deAnibal, Fernanda de Freitashttp://lattes.cnpq.br/4918261968772806http://lattes.cnpq.br/29161206549521833e66ce8b-852c-46a7-9b1a-9f350ac4ec7d2017-01-16T12:39:43Z2017-01-16T12:39:43Z2016-07-15SOUZA, Laís Cristina de. Reconhecimento da ligação dos anticorpos anti-HCV com proteínas recombinantes do vírus da hepatite C por meio do teste ELISA. 2016. Tese (Doutorado em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2016. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8396.https://repositorio.ufscar.br/handle/ufscar/8396Viral hepatitis is a major public health problem worldwide and in Brazil. They are notifiable diseases and according to estimates, billions of people have had contact with the hepatitis and millions are chronic carriers. Infection with hepatitis C virus (VHC) is a major problem worldwide public health due to the high rate of progression to chronicity, the evolutionary potential for cirrhosis and hepatocellular carcinoma, major complications leading to death. In general it can be said that in the last decade there have been major advances in the diagnosis of hepatitis C. In this period there was progressive improvement in sensitivity and specificity of the tests used to detect antibodies against the VHC virus. However, it is necessary that more accurate tests are developed. Thus, considering that the concern for the detection of hepatitis C increases every day, especially in blood banks; the diagnostic methods of this infection are of great clinical relevance and may be used as markers chronicity and indicative of therapeutic efficacy. Therefore, this work proposed to evaluate the connection and recognition of anti-VHC antibody positive and positive genotyped samples in patients with hepatitis C through the standardization of procedures and solutions used in qualitative ELISA. There was the process of awareness of microplates with recombinant chimeric protein, made the analysis of sensitivity, specificity, reproducibility and validity of the method. We obtained from ELISA assays with standardized recombinant proteins a protocol able to have a good performance of the main components of the reaction, and antigens conjugated with good resolution. This study presented ELISA results valid 95.69%, 100% reproducibility, 94.5% sensitivity and specificity 99.3%, higher than the ELISA performed with multiepitopo protein MEHCV who presented with sensitivity (92.86%) and specificity (82.89%). The standard ELISA can be used as a qualitative serological technique aimed at detection of anti-VHC antibodies, as demonstrated with great reactivity in patients infected with VHC.As hepatites virais são um grave problema de saúde pública no mundo e no Brasil. São doenças de notificação compulsória e segundo estimativas, bilhões de pessoas já tiveram contato com vírus das hepatites e milhões são portadores crônicos. A infecção pelo vírus da hepatite C (HCV) constitui um grave problema de saúde pública mundial devido à elevada taxa de progressão para cronicidade, ao potencial evolutivo para cirrose e carcinoma hepatocelular, principais complicações conducentes à morte. Em geral, pode-se dizer que na última década houve grandes avanços no diagnóstico da hepatite C. Nesse período houve progressiva melhora na sensibilidade e especificidade dos testes utilizados para detecção de anticorpos contra o vírus HCV. Contudo, é necessário que sejam desenvolvidos testes de maior acurácia. Assim, considerando que a preocupação com a detecção da hepatite C aumenta a cada dia, principalmente em bancos de sangue; os métodos diagnósticos desta infecção são de grande relevância clínica e podem ser utilizados como marcadores de cronicidade e indicativos da eficácia terapêutica. Portanto, esse trabalho propôs avaliar a ligação e reconhecimento dos anticorpos anti-HCV de amostras positivas e positivas genotipadas de pacientes portadores de Hepatite C, através da padronização dos procedimentos e soluções utilizadas no ELISA qualitativo. Realizou-se o processo de sensibilização das microplacas com proteína recombinante quimérica, fez-se a análise da sensibilidade, especificidade, reprodutibilidade e validade do método. Obtivemos a partir dos ensaios de padronização do ELISA com proteínas recombinantes um protocolo capaz de ter um bom rendimento dos principais componentes da reação, antígenos e conjugado, com boa resolução. O presente estudo apresentou-se resultados do ELISA com validade 95,69% , reprodutibilidade 100%, sensibilidade 94,5% e especificidade 99,3%, superior ao ELISA realizado com a proteína multiepitopo MEHCV que apresentaram com sensibilidade (92,86%) e especificidade (82,89%). O ELISA padronizado pode ser utilizado como uma técnica sorológica qualitativa, visando a detecção de anticorpos anti-HCV, pois mostrou-se com ótima reatividade nos soros pacientes infectados com HCV.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)porUniversidade Federal de São CarlosCâmpus São CarlosPrograma de Pós-Graduação em Biotecnologia - PPGBiotecUFSCarHepatitis CVHCELISA and recombinant proteinsProteínas recombinantesELISAOUTROSReconhecimento da ligação dos anticorpos anti-HCV com proteínas recombinantes do vírus da hepatite C por meio do teste ELISAinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisOnline600600d0b619ca-16cf-40f9-9e9b-1792083fa39finfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALTeseLCS.pdfTeseLCS.pdfapplication/pdf1420745https://repositorio.ufscar.br/bitstream/ufscar/8396/1/TeseLCS.pdf4dd4fbff68d9aa7235de8b5f959ba86dMD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81957https://repositorio.ufscar.br/bitstream/ufscar/8396/2/license.txtae0398b6f8b235e40ad82cba6c50031dMD52TEXTTeseLCS.pdf.txtTeseLCS.pdf.txtExtracted texttext/plain122431https://repositorio.ufscar.br/bitstream/ufscar/8396/3/TeseLCS.pdf.txtfa7b64e454da6e85cbad44fab3bc72ccMD53THUMBNAILTeseLCS.pdf.jpgTeseLCS.pdf.jpgIM Thumbnailimage/jpeg6524https://repositorio.ufscar.br/bitstream/ufscar/8396/4/TeseLCS.pdf.jpg1d76756949625283f9661554f099957eMD54ufscar/83962023-09-18 18:31:05.914oai:repositorio.ufscar.br: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Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:05Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Reconhecimento da ligação dos anticorpos anti-HCV com proteínas recombinantes do vírus da hepatite C por meio do teste ELISA |
title |
Reconhecimento da ligação dos anticorpos anti-HCV com proteínas recombinantes do vírus da hepatite C por meio do teste ELISA |
spellingShingle |
Reconhecimento da ligação dos anticorpos anti-HCV com proteínas recombinantes do vírus da hepatite C por meio do teste ELISA Souza, Laís Cristina de Hepatitis C VHC ELISA and recombinant proteins Proteínas recombinantes ELISA OUTROS |
title_short |
Reconhecimento da ligação dos anticorpos anti-HCV com proteínas recombinantes do vírus da hepatite C por meio do teste ELISA |
title_full |
Reconhecimento da ligação dos anticorpos anti-HCV com proteínas recombinantes do vírus da hepatite C por meio do teste ELISA |
title_fullStr |
Reconhecimento da ligação dos anticorpos anti-HCV com proteínas recombinantes do vírus da hepatite C por meio do teste ELISA |
title_full_unstemmed |
Reconhecimento da ligação dos anticorpos anti-HCV com proteínas recombinantes do vírus da hepatite C por meio do teste ELISA |
title_sort |
Reconhecimento da ligação dos anticorpos anti-HCV com proteínas recombinantes do vírus da hepatite C por meio do teste ELISA |
author |
Souza, Laís Cristina de |
author_facet |
Souza, Laís Cristina de |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/2916120654952183 |
dc.contributor.author.fl_str_mv |
Souza, Laís Cristina de |
dc.contributor.advisor1.fl_str_mv |
Anibal, Fernanda de Freitas |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/4918261968772806 |
dc.contributor.authorID.fl_str_mv |
3e66ce8b-852c-46a7-9b1a-9f350ac4ec7d |
contributor_str_mv |
Anibal, Fernanda de Freitas |
dc.subject.eng.fl_str_mv |
Hepatitis C VHC ELISA and recombinant proteins Proteínas recombinantes ELISA |
topic |
Hepatitis C VHC ELISA and recombinant proteins Proteínas recombinantes ELISA OUTROS |
dc.subject.cnpq.fl_str_mv |
OUTROS |
description |
Viral hepatitis is a major public health problem worldwide and in Brazil. They are notifiable diseases and according to estimates, billions of people have had contact with the hepatitis and millions are chronic carriers. Infection with hepatitis C virus (VHC) is a major problem worldwide public health due to the high rate of progression to chronicity, the evolutionary potential for cirrhosis and hepatocellular carcinoma, major complications leading to death. In general it can be said that in the last decade there have been major advances in the diagnosis of hepatitis C. In this period there was progressive improvement in sensitivity and specificity of the tests used to detect antibodies against the VHC virus. However, it is necessary that more accurate tests are developed. Thus, considering that the concern for the detection of hepatitis C increases every day, especially in blood banks; the diagnostic methods of this infection are of great clinical relevance and may be used as markers chronicity and indicative of therapeutic efficacy. Therefore, this work proposed to evaluate the connection and recognition of anti-VHC antibody positive and positive genotyped samples in patients with hepatitis C through the standardization of procedures and solutions used in qualitative ELISA. There was the process of awareness of microplates with recombinant chimeric protein, made the analysis of sensitivity, specificity, reproducibility and validity of the method. We obtained from ELISA assays with standardized recombinant proteins a protocol able to have a good performance of the main components of the reaction, and antigens conjugated with good resolution. This study presented ELISA results valid 95.69%, 100% reproducibility, 94.5% sensitivity and specificity 99.3%, higher than the ELISA performed with multiepitopo protein MEHCV who presented with sensitivity (92.86%) and specificity (82.89%). The standard ELISA can be used as a qualitative serological technique aimed at detection of anti-VHC antibodies, as demonstrated with great reactivity in patients infected with VHC. |
publishDate |
2016 |
dc.date.issued.fl_str_mv |
2016-07-15 |
dc.date.accessioned.fl_str_mv |
2017-01-16T12:39:43Z |
dc.date.available.fl_str_mv |
2017-01-16T12:39:43Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
SOUZA, Laís Cristina de. Reconhecimento da ligação dos anticorpos anti-HCV com proteínas recombinantes do vírus da hepatite C por meio do teste ELISA. 2016. Tese (Doutorado em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2016. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8396. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/8396 |
identifier_str_mv |
SOUZA, Laís Cristina de. Reconhecimento da ligação dos anticorpos anti-HCV com proteínas recombinantes do vírus da hepatite C por meio do teste ELISA. 2016. Tese (Doutorado em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2016. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8396. |
url |
https://repositorio.ufscar.br/handle/ufscar/8396 |
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Universidade Federal de São Carlos Câmpus São Carlos |
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Programa de Pós-Graduação em Biotecnologia - PPGBiotec |
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UFSCar |
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Universidade Federal de São Carlos Câmpus São Carlos |
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