Desenvolvimento de imunossensores microfluídicos descartáveis para detecção de biomarcadores visando o diagnóstico precoce da doença de Alzheimer
Autor(a) principal: | |
---|---|
Data de Publicação: | 2019 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/11546 |
Resumo: | Alzheimer's disease (AD) is a neurodegenerative condition that affects a large number of elderly and has an important social and economic impact. In early stages, the AD diagnosis can significantly improve patients' quality of life. Currently, AD biomarkers require diagnostic imaging procedures that are expensive or invasive. Thus, methods less invasive that allow an early, fast, and low-cost diagnosis are highly desirable. For this reason, a disposable microfluidic device was developed based on electrochemical immunosensors for detection of the biomarkers A Disintegrin And Metalloprotease 10 (ADAM10) and amyloid-β peptide with 42 amino acids (Aβ42) in plasma and cerebrospinal fluid samples. Three elderly groups divided into cognitively healthy subjects, mild cognitive impairment (MCI), and AD patients at different disease stages were analyzed. For device construction, electrodes were screen printed on polyester foil using carbon and Ag/AgCl inks. Strategies for the biomarker’s detection were based on sandwich or competitive immunoassays. The horseradish peroxidase (HRP) enzyme was used as an electrochemical marker and responsible for the indirect response of the immunosensor. The ADAM10 calibration curve was linear in the range of 5.6 to 1.4 pg mL-1, with a linear correlation coefficient (r2) of 0.983, and a limit of detection (LD) of 5.6 fg mL-1. For βA42 biomarker, the concentration range was linear between 1.8 and 499.2 pg mL-1 of the peptide, with r2 = 0.977 and LD of 1.8 pg mL-1. Plasma samples from AD and MCI patients showed significantly increased of ADAM10 levels when compared with healthy subjects. Aβ42 concentrations found in the cerebrospinal fluid were statistically different between the control and the diseased groups. ADAM10 and its detection using the device proposed here proved to be a powerful tool for the early diagnosis and AD monitoring, which can bring great benefits to patients' quality of life. |
id |
SCAR_efb2ec1c0dd46c3fc3495d32e808a60a |
---|---|
oai_identifier_str |
oai:repositorio.ufscar.br:ufscar/11546 |
network_acronym_str |
SCAR |
network_name_str |
Repositório Institucional da UFSCAR |
repository_id_str |
4322 |
spelling |
Oliveira, Tássia Regina deFaria, Ronaldo Censihttp://lattes.cnpq.br/8659496864305621http://lattes.cnpq.br/9671451657511244286e9f4e-a1da-4615-9750-14626a7fec302019-07-19T18:12:55Z2019-07-19T18:12:55Z2019-03-25OLIVEIRA, Tássia Regina de. Desenvolvimento de imunossensores microfluídicos descartáveis para detecção de biomarcadores visando o diagnóstico precoce da doença de Alzheimer. 2019. Tese (Doutorado em Química) – Universidade Federal de São Carlos, São Carlos, 2019. Disponível em: https://repositorio.ufscar.br/handle/ufscar/11546.https://repositorio.ufscar.br/handle/ufscar/11546Alzheimer's disease (AD) is a neurodegenerative condition that affects a large number of elderly and has an important social and economic impact. In early stages, the AD diagnosis can significantly improve patients' quality of life. Currently, AD biomarkers require diagnostic imaging procedures that are expensive or invasive. Thus, methods less invasive that allow an early, fast, and low-cost diagnosis are highly desirable. For this reason, a disposable microfluidic device was developed based on electrochemical immunosensors for detection of the biomarkers A Disintegrin And Metalloprotease 10 (ADAM10) and amyloid-β peptide with 42 amino acids (Aβ42) in plasma and cerebrospinal fluid samples. Three elderly groups divided into cognitively healthy subjects, mild cognitive impairment (MCI), and AD patients at different disease stages were analyzed. For device construction, electrodes were screen printed on polyester foil using carbon and Ag/AgCl inks. Strategies for the biomarker’s detection were based on sandwich or competitive immunoassays. The horseradish peroxidase (HRP) enzyme was used as an electrochemical marker and responsible for the indirect response of the immunosensor. The ADAM10 calibration curve was linear in the range of 5.6 to 1.4 pg mL-1, with a linear correlation coefficient (r2) of 0.983, and a limit of detection (LD) of 5.6 fg mL-1. For βA42 biomarker, the concentration range was linear between 1.8 and 499.2 pg mL-1 of the peptide, with r2 = 0.977 and LD of 1.8 pg mL-1. Plasma samples from AD and MCI patients showed significantly increased of ADAM10 levels when compared with healthy subjects. Aβ42 concentrations found in the cerebrospinal fluid were statistically different between the control and the diseased groups. ADAM10 and its detection using the device proposed here proved to be a powerful tool for the early diagnosis and AD monitoring, which can bring great benefits to patients' quality of life.A doença de Alzheimer (DA) é uma doença neurodegenerativa que afeta um grande número de idosos e tem grande impacto social e econômico. Em estágios iniciais, o diagnóstico da DA pode melhorar significativamente a qualidade de vida dos pacientes. Atualmente, os biomarcadores da DA requerem procedimentos de diagnóstico de imagem dispendiosos ou invasivos. Desta maneira, métodos que permitem um diagnóstico precoce, menos invasivo, rápido e de baixo custo são altamente desejáveis. Por esse motivo, foi desenvolvido um dispositivo microfluídico descartável baseado em imunossensores eletroquímicos para a detecção dos biomarcadores A Disintegrin And Metalloprotease 10 (ADAM10) em plasma e peptídeo β-amiloide com 42 aminoácidos (βA42) em liquor. Amostras de três grupos de idosos divididos em sujeitos cognitivamente saudáveis, pacientes com transtorno neurocognitivo leve (TNCL) e com DA em diferentes estágios foram analisadas. Para a construção do dispositivo, eletrodos foram serigrafados em folha de poliéster utilizando tintas à base de carbono e Ag/AgCl. As estratégias para detecção dos biomarcadores foram baseadas nos imunoensaios do tipo sanduíche ou competitivo indireto. A enzima peroxidase de raiz forte (HRP) foi utilizada como marcador eletroquímico e responsável pela resposta indireta do imunossensor. A curva analítica para a ADAM10 foi linear na faixa de 5,6 a 1,4 pg mL-1, com coeficiente de correlação linear (r2) de 0,983 e limite de detecção (LD) de 5,6 fg mL-1. Para o biomarcador βA42, a faixa foi linear entre 1,8 a 499,2 pg mL-1 do peptídeo, com r2 = 0,977 e LD de 1,8 pg mL-1. Amostras de plasma dos pacientes com DA e TNCL apresentaram níveis de ADAM10 significativamente aumentados quando comparado com sujeitos saudáveis. As concentrações de βA42 encontradas no liquor foram estatisticamente diferentes entre os grupos controle e doentes. A ADAM10 e sua detecção utilizando o dispositivo aqui proposto mostraram-se como uma poderosa ferramenta para o diagnóstico precoce e monitoramento da DA o que pode trazer grandes benefícios para a qualidade de vida dos pacientes.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP: 2014/22401-0porUniversidade Federal de São CarlosCâmpus São CarlosPrograma de Pós-Graduação em Química - PPGQUFSCarAlzheimer's diseaseImmunosensorMicrofluidic deviceEarly diagnosisADAM10βA42CIENCIAS EXATAS E DA TERRA::QUIMICA::QUIMICA ANALITICA::ELETROANALITICADesenvolvimento de imunossensores microfluídicos descartáveis para detecção de biomarcadores visando o diagnóstico precoce da doença de AlzheimerDevelopment of disposable microfluidic immunosensors for biomarker detection aiming early diagnosis of Alzheimer's diseaseinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisOnline0e05d8c5-0f80-49a6-98a8-b6645fe6f1e3info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALTESE TÁSSIA REGINA DE OLIVEIRA.pdfTESE TÁSSIA REGINA DE OLIVEIRA.pdfTese Tássia Regina de Oliveiraapplication/pdf3871517https://repositorio.ufscar.br/bitstream/ufscar/11546/5/TESE%20T%c3%81SSIA%20REGINA%20DE%20OLIVEIRA.pdfe0382e334443f7b0255a567304d112a0MD55LICENSElicense.txtlicense.txttext/plain; charset=utf-81957https://repositorio.ufscar.br/bitstream/ufscar/11546/6/license.txtae0398b6f8b235e40ad82cba6c50031dMD56TEXTTESE TÁSSIA REGINA DE OLIVEIRA.pdf.txtTESE TÁSSIA REGINA DE OLIVEIRA.pdf.txtExtracted texttext/plain186282https://repositorio.ufscar.br/bitstream/ufscar/11546/7/TESE%20T%c3%81SSIA%20REGINA%20DE%20OLIVEIRA.pdf.txtc8c00b1b96223d3d63d0675d151f796cMD57THUMBNAILTESE TÁSSIA REGINA DE OLIVEIRA.pdf.jpgTESE TÁSSIA REGINA DE OLIVEIRA.pdf.jpgIM Thumbnailimage/jpeg9798https://repositorio.ufscar.br/bitstream/ufscar/11546/8/TESE%20T%c3%81SSIA%20REGINA%20DE%20OLIVEIRA.pdf.jpgf0479264d6f09782e01a51d351950b0fMD58ufscar/115462023-09-18 18:31:13.628oai:repositorio.ufscar.br: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Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:13Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Desenvolvimento de imunossensores microfluídicos descartáveis para detecção de biomarcadores visando o diagnóstico precoce da doença de Alzheimer |
dc.title.alternative.eng.fl_str_mv |
Development of disposable microfluidic immunosensors for biomarker detection aiming early diagnosis of Alzheimer's disease |
title |
Desenvolvimento de imunossensores microfluídicos descartáveis para detecção de biomarcadores visando o diagnóstico precoce da doença de Alzheimer |
spellingShingle |
Desenvolvimento de imunossensores microfluídicos descartáveis para detecção de biomarcadores visando o diagnóstico precoce da doença de Alzheimer Oliveira, Tássia Regina de Alzheimer's disease Immunosensor Microfluidic device Early diagnosis ADAM10 βA42 CIENCIAS EXATAS E DA TERRA::QUIMICA::QUIMICA ANALITICA::ELETROANALITICA |
title_short |
Desenvolvimento de imunossensores microfluídicos descartáveis para detecção de biomarcadores visando o diagnóstico precoce da doença de Alzheimer |
title_full |
Desenvolvimento de imunossensores microfluídicos descartáveis para detecção de biomarcadores visando o diagnóstico precoce da doença de Alzheimer |
title_fullStr |
Desenvolvimento de imunossensores microfluídicos descartáveis para detecção de biomarcadores visando o diagnóstico precoce da doença de Alzheimer |
title_full_unstemmed |
Desenvolvimento de imunossensores microfluídicos descartáveis para detecção de biomarcadores visando o diagnóstico precoce da doença de Alzheimer |
title_sort |
Desenvolvimento de imunossensores microfluídicos descartáveis para detecção de biomarcadores visando o diagnóstico precoce da doença de Alzheimer |
author |
Oliveira, Tássia Regina de |
author_facet |
Oliveira, Tássia Regina de |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/9671451657511244 |
dc.contributor.author.fl_str_mv |
Oliveira, Tássia Regina de |
dc.contributor.advisor1.fl_str_mv |
Faria, Ronaldo Censi |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/8659496864305621 |
dc.contributor.authorID.fl_str_mv |
286e9f4e-a1da-4615-9750-14626a7fec30 |
contributor_str_mv |
Faria, Ronaldo Censi |
dc.subject.eng.fl_str_mv |
Alzheimer's disease Immunosensor Microfluidic device Early diagnosis ADAM10 βA42 |
topic |
Alzheimer's disease Immunosensor Microfluidic device Early diagnosis ADAM10 βA42 CIENCIAS EXATAS E DA TERRA::QUIMICA::QUIMICA ANALITICA::ELETROANALITICA |
dc.subject.cnpq.fl_str_mv |
CIENCIAS EXATAS E DA TERRA::QUIMICA::QUIMICA ANALITICA::ELETROANALITICA |
description |
Alzheimer's disease (AD) is a neurodegenerative condition that affects a large number of elderly and has an important social and economic impact. In early stages, the AD diagnosis can significantly improve patients' quality of life. Currently, AD biomarkers require diagnostic imaging procedures that are expensive or invasive. Thus, methods less invasive that allow an early, fast, and low-cost diagnosis are highly desirable. For this reason, a disposable microfluidic device was developed based on electrochemical immunosensors for detection of the biomarkers A Disintegrin And Metalloprotease 10 (ADAM10) and amyloid-β peptide with 42 amino acids (Aβ42) in plasma and cerebrospinal fluid samples. Three elderly groups divided into cognitively healthy subjects, mild cognitive impairment (MCI), and AD patients at different disease stages were analyzed. For device construction, electrodes were screen printed on polyester foil using carbon and Ag/AgCl inks. Strategies for the biomarker’s detection were based on sandwich or competitive immunoassays. The horseradish peroxidase (HRP) enzyme was used as an electrochemical marker and responsible for the indirect response of the immunosensor. The ADAM10 calibration curve was linear in the range of 5.6 to 1.4 pg mL-1, with a linear correlation coefficient (r2) of 0.983, and a limit of detection (LD) of 5.6 fg mL-1. For βA42 biomarker, the concentration range was linear between 1.8 and 499.2 pg mL-1 of the peptide, with r2 = 0.977 and LD of 1.8 pg mL-1. Plasma samples from AD and MCI patients showed significantly increased of ADAM10 levels when compared with healthy subjects. Aβ42 concentrations found in the cerebrospinal fluid were statistically different between the control and the diseased groups. ADAM10 and its detection using the device proposed here proved to be a powerful tool for the early diagnosis and AD monitoring, which can bring great benefits to patients' quality of life. |
publishDate |
2019 |
dc.date.accessioned.fl_str_mv |
2019-07-19T18:12:55Z |
dc.date.available.fl_str_mv |
2019-07-19T18:12:55Z |
dc.date.issued.fl_str_mv |
2019-03-25 |
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 |
OLIVEIRA, Tássia Regina de. Desenvolvimento de imunossensores microfluídicos descartáveis para detecção de biomarcadores visando o diagnóstico precoce da doença de Alzheimer. 2019. Tese (Doutorado em Química) – Universidade Federal de São Carlos, São Carlos, 2019. Disponível em: https://repositorio.ufscar.br/handle/ufscar/11546. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/11546 |
identifier_str_mv |
OLIVEIRA, Tássia Regina de. Desenvolvimento de imunossensores microfluídicos descartáveis para detecção de biomarcadores visando o diagnóstico precoce da doença de Alzheimer. 2019. Tese (Doutorado em Química) – Universidade Federal de São Carlos, São Carlos, 2019. Disponível em: https://repositorio.ufscar.br/handle/ufscar/11546. |
url |
https://repositorio.ufscar.br/handle/ufscar/11546 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.authority.fl_str_mv |
0e05d8c5-0f80-49a6-98a8-b6645fe6f1e3 |
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 Química - PPGQ |
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/11546/5/TESE%20T%c3%81SSIA%20REGINA%20DE%20OLIVEIRA.pdf https://repositorio.ufscar.br/bitstream/ufscar/11546/6/license.txt https://repositorio.ufscar.br/bitstream/ufscar/11546/7/TESE%20T%c3%81SSIA%20REGINA%20DE%20OLIVEIRA.pdf.txt https://repositorio.ufscar.br/bitstream/ufscar/11546/8/TESE%20T%c3%81SSIA%20REGINA%20DE%20OLIVEIRA.pdf.jpg |
bitstream.checksum.fl_str_mv |
e0382e334443f7b0255a567304d112a0 ae0398b6f8b235e40ad82cba6c50031d c8c00b1b96223d3d63d0675d151f796c f0479264d6f09782e01a51d351950b0f |
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_ |
1802136359979188224 |