Reconhecimento da ligação dos anticorpos anti-HCV com proteínas recombinantes do vírus da hepatite C por meio do teste ELISA

Detalhes bibliográficos
Ano de defesa: 2016
Autor(a) principal: Souza, Laís Cristina de
Orientador(a): Anibal, Fernanda de Freitas lattes
Banca de defesa: Não Informado pela instituição
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:
VHC
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.
id SCAR_f5ff57c8370730007a3501fefafc96d3
oai_identifier_str oai:repositorio.ufscar.br:ufscar/8396
network_acronym_str SCAR
network_name_str Repositório Institucional da UFSCAR
repository_id_str
spelling 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
status_str 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
dc.language.iso.fl_str_mv por
language por
dc.relation.confidence.fl_str_mv 600
600
dc.relation.authority.fl_str_mv d0b619ca-16cf-40f9-9e9b-1792083fa39f
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Universidade Federal de São Carlos
Câmpus São Carlos
dc.publisher.program.fl_str_mv Programa de Pós-Graduação em Biotecnologia - PPGBiotec
dc.publisher.initials.fl_str_mv UFSCar
publisher.none.fl_str_mv Universidade Federal de São Carlos
Câmpus São Carlos
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFSCAR
instname:Universidade Federal de São Carlos (UFSCAR)
instacron:UFSCAR
instname_str Universidade Federal de São Carlos (UFSCAR)
instacron_str UFSCAR
institution UFSCAR
reponame_str Repositório Institucional da UFSCAR
collection Repositório Institucional da UFSCAR
bitstream.url.fl_str_mv https://repositorio.ufscar.br/bitstream/ufscar/8396/1/TeseLCS.pdf
https://repositorio.ufscar.br/bitstream/ufscar/8396/2/license.txt
https://repositorio.ufscar.br/bitstream/ufscar/8396/3/TeseLCS.pdf.txt
https://repositorio.ufscar.br/bitstream/ufscar/8396/4/TeseLCS.pdf.jpg
bitstream.checksum.fl_str_mv 4dd4fbff68d9aa7235de8b5f959ba86d
ae0398b6f8b235e40ad82cba6c50031d
fa7b64e454da6e85cbad44fab3bc72cc
1d76756949625283f9661554f099957e
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
MD5
MD5
repository.name.fl_str_mv Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)
repository.mail.fl_str_mv
_version_ 1802136525719207936