Dispositivos microfluídicos baseados em fita adesiva dupla face e aplicações no diagnóstico molecular da Covid-19
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFG |
Texto Completo: | http://repositorio.bc.ufg.br/tede/handle/tede/11612 |
Resumo: | Microfluidic devices have shown potential application in several areas of science due to the possibility of integration and automation of steps, reducing time and cost in total analysis. However, due to the laboriousness of the preparation, its use is still discreet. Thus, in the aim to make microfluidic devices accessible, in this study we explore the development of disposable microfluidic devices based on double-sided tape and polyester film (DF-Pe) and we initially demonstrated its use in all stages of molecular diagnosis for COVID-19. Devices based on double-sided adhesive tape were manufactured in two different forms: using a laser cutter or totally free-instrumentation. The DF-Pe microchips developed here exhibited low cost, simplicity, fast fabrication, chemical resistance, versatility for layout changes and biocompatibility. In the DF-Pe microchips, we carried out the isolation of nucleic acids from samples of patients with SARS-CoV-2 infection, using the technique of dynamic solid phase extraction (dSPE) with an efficiency of extraction of 56%, obtaining the RNA at the end of the process suitable to be amplified by RT-qPCR (Reverse Transcription - Quantitative Pol-ymerase Chain Reaction) and RT-LAMP (Reverse Transcription - Loop-Mediated Isother-mal Amplification). We demonstrated a RT-LAMP based assay to detect SARS-CoV-2 di-rectly on microdevices in 30 minutes, also we applied the test to a panel of 40 samples and the results obtained were with 100% concordance with the gold standard method (RT-qPCR). Furthermore, the features of the DF-Pe microdevice were evaluated through the development of passive micromixers that allowed visual detection of the amplification prod-ucts directly on the microdevices. The high simplicity, cost-effective and fast manufacturing of the DF-Pe microdevices, integrated to the simple and quick extraction, amplification and viral detection directly on chip, demonstrates the high potential application of this method in the molecular diagnosis of COVID-19 in resources limited settings. |
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Duarte, Gabriela Rodrigues Mendeshttp://lattes.cnpq.br/9005971441891787Duarte, Gabriela Rodrigues MendesGomes, Danielle Cangussu de CastroRodrigues, Flávia Melohttp://lattes.cnpq.br/2591378387192528Estrela, Paulo Felipe Neves2021-08-31T11:20:34Z2021-08-31T11:20:34Z2021-07-29ESTRELA, P. F. N. Dispositivos microfluídicos baseados em fita adesiva dupla face e aplicações no diagnóstico molecular da Covid-19. 2021. 93 f. Dissertação (Mestrado em Química) - Universidade Federal de Goiás, Goiânia, 2021.http://repositorio.bc.ufg.br/tede/handle/tede/11612ark:/38995/001300000c0vmMicrofluidic devices have shown potential application in several areas of science due to the possibility of integration and automation of steps, reducing time and cost in total analysis. However, due to the laboriousness of the preparation, its use is still discreet. Thus, in the aim to make microfluidic devices accessible, in this study we explore the development of disposable microfluidic devices based on double-sided tape and polyester film (DF-Pe) and we initially demonstrated its use in all stages of molecular diagnosis for COVID-19. Devices based on double-sided adhesive tape were manufactured in two different forms: using a laser cutter or totally free-instrumentation. The DF-Pe microchips developed here exhibited low cost, simplicity, fast fabrication, chemical resistance, versatility for layout changes and biocompatibility. In the DF-Pe microchips, we carried out the isolation of nucleic acids from samples of patients with SARS-CoV-2 infection, using the technique of dynamic solid phase extraction (dSPE) with an efficiency of extraction of 56%, obtaining the RNA at the end of the process suitable to be amplified by RT-qPCR (Reverse Transcription - Quantitative Pol-ymerase Chain Reaction) and RT-LAMP (Reverse Transcription - Loop-Mediated Isother-mal Amplification). We demonstrated a RT-LAMP based assay to detect SARS-CoV-2 di-rectly on microdevices in 30 minutes, also we applied the test to a panel of 40 samples and the results obtained were with 100% concordance with the gold standard method (RT-qPCR). Furthermore, the features of the DF-Pe microdevice were evaluated through the development of passive micromixers that allowed visual detection of the amplification prod-ucts directly on the microdevices. The high simplicity, cost-effective and fast manufacturing of the DF-Pe microdevices, integrated to the simple and quick extraction, amplification and viral detection directly on chip, demonstrates the high potential application of this method in the molecular diagnosis of COVID-19 in resources limited settings.Dispositivos microfluídicos têm demonstrado potencial para aplicações em diversos ramos da ciência pela possibilidade de integração e automatização de etapas, redução do tempo e custos em análises totais. Entretanto, devido a laboriosidade de confecção, sua utilização ainda é discreta. Nesta perspectiva, visando tornar os dispositivos microfluídicos mais acessíveis, exploramos neste trabalho o desenvolvimento de microdispositivos descartáveis à base de fita adesiva dupla face e filme poliéster (DF-Pe) e demonstramos, inicialmente, sua utilização em todas as etapas do diagnóstico molecular para COVID-19. Dispositivos a base de fita adesiva dupla face foram fabricados de dois modos distintos: utilizando uma cortadora a laser ou fabricados totalmente livres de instrumentação. Os microchips DF-Pe desenvolvidos apresentaram baixo custo, simplicidade e rapidez para confecção, resistência química, versatilidade para mudanças de layouts e biocompatibilidade. Realizamos nos microchips DF-Pe o isolamento de ácidos nucleicos de amostras de pacientes com infecção por SARS-CoV-2, por meio da técnica de extração dinâmica em fase sólida (dSPE), com eficiência média de extração de 56%, obtendo ao final do processo RNA suficientemente íntegro para ser amplificado por RT-qPCR (Transcrição Reversa seguida da Reação em Cadeia da Polimerase quantitativa) e RT-LAMP (Transcrição Reversa seguida da Amplificação Isotérmica Mediada por Loop). Demonstramos a RT-LAMP para detecção de SARS-CoV-2 diretamente nos microdispositivos em 30 minutos e, ao aplicarmos o teste a um painel de 40 amostras, obtivemos resultados 100% concordantes com método padrão ouro (RT-qPCR). Ademais, as funcionalidades dos microdispositivos DF-Pe foram exploradas no desenvolvimento de micromisturadores passivos que permitiram detecção visual de produtos de amplificação diretamente nos microdispositivos. A elevada simplicidade, rapidez e baixo custo na fabricação dos microdispositivos DF-Pe, associada à simples e rápida extração, amplificação e detecção viral on chip, demonstra o elevado potencial de aplicação dos microdispositivos DF-Pe para o diagnóstico molecular da COVID-19 em locais que detém poucos recursos.Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2021-08-31T03:21:02Z No. of bitstreams: 2 Dissertação - Paulo Felipe Neves Estrela - 2021.pdf: 6854103 bytes, checksum: 1120eb56a0efc72ec25cd0eb120e1b77 (MD5) license_rdf: 805 bytes, checksum: 4460e5956bc1d1639be9ae6146a50347 (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2021-08-31T11:20:34Z (GMT) No. of bitstreams: 2 Dissertação - Paulo Felipe Neves Estrela - 2021.pdf: 6854103 bytes, checksum: 1120eb56a0efc72ec25cd0eb120e1b77 (MD5) license_rdf: 805 bytes, checksum: 4460e5956bc1d1639be9ae6146a50347 (MD5)Made available in DSpace on 2021-08-31T11:20:34Z (GMT). No. of bitstreams: 2 Dissertação - Paulo Felipe Neves Estrela - 2021.pdf: 6854103 bytes, checksum: 1120eb56a0efc72ec25cd0eb120e1b77 (MD5) license_rdf: 805 bytes, checksum: 4460e5956bc1d1639be9ae6146a50347 (MD5) Previous issue date: 2021-07-29Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPqFundação de Amparo à Pesquisa do Estado de GoiásOutroporUniversidade Federal de GoiásPrograma de Pós-graduação em Química (IQ)UFGBrasilInstituto de Química - IQ (RG)Attribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessMicrochipSARS-CoV-2BioanalíticaLoop-mediated isothermal amplificationBioanalyticsCIENCIAS EXATAS E DA TERRA::QUIMICA::QUIMICA-ANALITICADispositivos microfluídicos baseados em fita adesiva dupla face e aplicações no diagnóstico molecular da Covid-19Microfluidic devices based on double-sided adhesive tape and applications in molecular diagnosis of Covid-19info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis82500500500500500500291092035reponame:Repositório Institucional da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGLICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.bc.ufg.br/tede/bitstreams/41adada0-9630-44ad-8c3a-807b1e9a4a09/download8a4605be74aa9ea9d79846c1fba20a33MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8805http://repositorio.bc.ufg.br/tede/bitstreams/a808c32b-d5f2-439f-964a-a9e33d50dcb6/download4460e5956bc1d1639be9ae6146a50347MD52ORIGINALDissertação - Paulo Felipe Neves Estrela - 2021.pdfDissertação - Paulo Felipe Neves Estrela - 2021.pdfapplication/pdf6854103http://repositorio.bc.ufg.br/tede/bitstreams/b597d124-4bb7-41fc-93da-eb15a37340a5/download1120eb56a0efc72ec25cd0eb120e1b77MD53tede/116122021-10-29 18:42:31.832http://creativecommons.org/licenses/by-nc-nd/4.0/Attribution-NonCommercial-NoDerivatives 4.0 Internationalopen.accessoai:repositorio.bc.ufg.br:tede/11612http://repositorio.bc.ufg.br/tedeRepositório InstitucionalPUBhttp://repositorio.bc.ufg.br/oai/requesttasesdissertacoes.bc@ufg.bropendoar:2021-10-29T21:42:31Repositório Institucional da UFG - Universidade Federal de Goiás (UFG)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 |
dc.title.pt_BR.fl_str_mv |
Dispositivos microfluídicos baseados em fita adesiva dupla face e aplicações no diagnóstico molecular da Covid-19 |
dc.title.alternative.eng.fl_str_mv |
Microfluidic devices based on double-sided adhesive tape and applications in molecular diagnosis of Covid-19 |
title |
Dispositivos microfluídicos baseados em fita adesiva dupla face e aplicações no diagnóstico molecular da Covid-19 |
spellingShingle |
Dispositivos microfluídicos baseados em fita adesiva dupla face e aplicações no diagnóstico molecular da Covid-19 Estrela, Paulo Felipe Neves Microchip SARS-CoV-2 Bioanalítica Loop-mediated isothermal amplification Bioanalytics CIENCIAS EXATAS E DA TERRA::QUIMICA::QUIMICA-ANALITICA |
title_short |
Dispositivos microfluídicos baseados em fita adesiva dupla face e aplicações no diagnóstico molecular da Covid-19 |
title_full |
Dispositivos microfluídicos baseados em fita adesiva dupla face e aplicações no diagnóstico molecular da Covid-19 |
title_fullStr |
Dispositivos microfluídicos baseados em fita adesiva dupla face e aplicações no diagnóstico molecular da Covid-19 |
title_full_unstemmed |
Dispositivos microfluídicos baseados em fita adesiva dupla face e aplicações no diagnóstico molecular da Covid-19 |
title_sort |
Dispositivos microfluídicos baseados em fita adesiva dupla face e aplicações no diagnóstico molecular da Covid-19 |
author |
Estrela, Paulo Felipe Neves |
author_facet |
Estrela, Paulo Felipe Neves |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Duarte, Gabriela Rodrigues Mendes |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/9005971441891787 |
dc.contributor.referee1.fl_str_mv |
Duarte, Gabriela Rodrigues Mendes |
dc.contributor.referee2.fl_str_mv |
Gomes, Danielle Cangussu de Castro |
dc.contributor.referee3.fl_str_mv |
Rodrigues, Flávia Melo |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/2591378387192528 |
dc.contributor.author.fl_str_mv |
Estrela, Paulo Felipe Neves |
contributor_str_mv |
Duarte, Gabriela Rodrigues Mendes Duarte, Gabriela Rodrigues Mendes Gomes, Danielle Cangussu de Castro Rodrigues, Flávia Melo |
dc.subject.por.fl_str_mv |
Microchip SARS-CoV-2 Bioanalítica |
topic |
Microchip SARS-CoV-2 Bioanalítica Loop-mediated isothermal amplification Bioanalytics CIENCIAS EXATAS E DA TERRA::QUIMICA::QUIMICA-ANALITICA |
dc.subject.eng.fl_str_mv |
Loop-mediated isothermal amplification Bioanalytics |
dc.subject.cnpq.fl_str_mv |
CIENCIAS EXATAS E DA TERRA::QUIMICA::QUIMICA-ANALITICA |
description |
Microfluidic devices have shown potential application in several areas of science due to the possibility of integration and automation of steps, reducing time and cost in total analysis. However, due to the laboriousness of the preparation, its use is still discreet. Thus, in the aim to make microfluidic devices accessible, in this study we explore the development of disposable microfluidic devices based on double-sided tape and polyester film (DF-Pe) and we initially demonstrated its use in all stages of molecular diagnosis for COVID-19. Devices based on double-sided adhesive tape were manufactured in two different forms: using a laser cutter or totally free-instrumentation. The DF-Pe microchips developed here exhibited low cost, simplicity, fast fabrication, chemical resistance, versatility for layout changes and biocompatibility. In the DF-Pe microchips, we carried out the isolation of nucleic acids from samples of patients with SARS-CoV-2 infection, using the technique of dynamic solid phase extraction (dSPE) with an efficiency of extraction of 56%, obtaining the RNA at the end of the process suitable to be amplified by RT-qPCR (Reverse Transcription - Quantitative Pol-ymerase Chain Reaction) and RT-LAMP (Reverse Transcription - Loop-Mediated Isother-mal Amplification). We demonstrated a RT-LAMP based assay to detect SARS-CoV-2 di-rectly on microdevices in 30 minutes, also we applied the test to a panel of 40 samples and the results obtained were with 100% concordance with the gold standard method (RT-qPCR). Furthermore, the features of the DF-Pe microdevice were evaluated through the development of passive micromixers that allowed visual detection of the amplification prod-ucts directly on the microdevices. The high simplicity, cost-effective and fast manufacturing of the DF-Pe microdevices, integrated to the simple and quick extraction, amplification and viral detection directly on chip, demonstrates the high potential application of this method in the molecular diagnosis of COVID-19 in resources limited settings. |
publishDate |
2021 |
dc.date.accessioned.fl_str_mv |
2021-08-31T11:20:34Z |
dc.date.available.fl_str_mv |
2021-08-31T11:20:34Z |
dc.date.issued.fl_str_mv |
2021-07-29 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
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masterThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
ESTRELA, P. F. N. Dispositivos microfluídicos baseados em fita adesiva dupla face e aplicações no diagnóstico molecular da Covid-19. 2021. 93 f. Dissertação (Mestrado em Química) - Universidade Federal de Goiás, Goiânia, 2021. |
dc.identifier.uri.fl_str_mv |
http://repositorio.bc.ufg.br/tede/handle/tede/11612 |
dc.identifier.dark.fl_str_mv |
ark:/38995/001300000c0vm |
identifier_str_mv |
ESTRELA, P. F. N. Dispositivos microfluídicos baseados em fita adesiva dupla face e aplicações no diagnóstico molecular da Covid-19. 2021. 93 f. Dissertação (Mestrado em Química) - Universidade Federal de Goiás, Goiânia, 2021. ark:/38995/001300000c0vm |
url |
http://repositorio.bc.ufg.br/tede/handle/tede/11612 |
dc.language.iso.fl_str_mv |
por |
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por |
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29 |
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0 3 5 |
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Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Goiás |
dc.publisher.program.fl_str_mv |
Programa de Pós-graduação em Química (IQ) |
dc.publisher.initials.fl_str_mv |
UFG |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Instituto de Química - IQ (RG) |
publisher.none.fl_str_mv |
Universidade Federal de Goiás |
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