Digital Microfluidics-Powered Real-Time Monitoring of Isothermal DNA Amplification of Cancer Biomarker

Bibliographic Details
Main Author: Coelho, Beatriz Jorge
Publication Date: 2022
Other Authors: Veigas, Bruno, Bettencourt, Luís, Águas, Hugo, Fortunato, Elvira, Martins, Rodrigo, Baptista, Pedro V., Igreja, Rui
Format: Article
Language: eng
Source: Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
Download full: http://hdl.handle.net/10362/142839
Summary: LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy?i4HB. Additional funding was received from FCT under the scope of the project reference LA/P/0037/2020, Beatriz J. Coelho and grant COVID/BD/152453/2022. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.
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spelling Digital Microfluidics-Powered Real-Time Monitoring of Isothermal DNA Amplification of Cancer Biomarkercancer biomarkerdigital microfluidicsfluorescence detectionloop-mediated isothermal amplificationreal-time nucleic acid amplification monitoringClinical BiochemistrySDG 3 - Good Health and Well-beingLA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy?i4HB. Additional funding was received from FCT under the scope of the project reference LA/P/0037/2020, Beatriz J. Coelho and grant COVID/BD/152453/2022. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.We introduce a digital microfluidics (DMF) platform specifically designed to perform a loop-mediated isothermal amplification (LAMP) of DNA and applied it to a real-time amplification to monitor a cancer biomarker, c-Myc (associated to 40% of all human tumors), using fluorescence microscopy. We demonstrate the full manipulation of the sample and reagents on the DMF platform, resulting in the successful amplification of 90 pg of the target DNA (0.5 ng/µL) in less than one hour. Furthermore, we test the efficiency of an innovative mixing strategy in DMF by employing two mixing methodologies onto the DMF droplets—low frequency AC (alternating current) actuation as well as back-and-forth droplet motion—which allows for improved fluorescence readouts. Fluo-rophore bleaching effects are minimized through on-chip sample partitioning by DMF processes and sequential droplet irradiation. Finally, LAMP reactions require only 2 µL volume droplets, which represents a 10-fold volume reduction in comparison to benchtop LAMP.DCM - Departamento de Ciência dos MateriaisUNINOVA-Instituto de Desenvolvimento de Novas TecnologiasDCV - Departamento de Ciências da VidaUCIBIO - Applied Molecular Biosciences UnitRUNCoelho, Beatriz JorgeVeigas, BrunoBettencourt, LuísÁguas, HugoFortunato, ElviraMartins, RodrigoBaptista, Pedro V.Igreja, Rui2022-08-03T22:26:02Z2022-03-282022-03-28T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article12application/pdfhttp://hdl.handle.net/10362/142839eng2079-6374PURE: 43542197https://doi.org/10.3390/bios12040201info:eu-repo/semantics/openAccessreponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiainstacron:RCAAP2024-05-22T18:04:16Zoai:run.unl.pt:10362/142839Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireinfo@rcaap.ptopendoar:https://opendoar.ac.uk/repository/71602025-05-28T17:34:47.376828Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiafalse
dc.title.none.fl_str_mv Digital Microfluidics-Powered Real-Time Monitoring of Isothermal DNA Amplification of Cancer Biomarker
title Digital Microfluidics-Powered Real-Time Monitoring of Isothermal DNA Amplification of Cancer Biomarker
spellingShingle Digital Microfluidics-Powered Real-Time Monitoring of Isothermal DNA Amplification of Cancer Biomarker
Coelho, Beatriz Jorge
cancer biomarker
digital microfluidics
fluorescence detection
loop-mediated isothermal amplification
real-time nucleic acid amplification monitoring
Clinical Biochemistry
SDG 3 - Good Health and Well-being
title_short Digital Microfluidics-Powered Real-Time Monitoring of Isothermal DNA Amplification of Cancer Biomarker
title_full Digital Microfluidics-Powered Real-Time Monitoring of Isothermal DNA Amplification of Cancer Biomarker
title_fullStr Digital Microfluidics-Powered Real-Time Monitoring of Isothermal DNA Amplification of Cancer Biomarker
title_full_unstemmed Digital Microfluidics-Powered Real-Time Monitoring of Isothermal DNA Amplification of Cancer Biomarker
title_sort Digital Microfluidics-Powered Real-Time Monitoring of Isothermal DNA Amplification of Cancer Biomarker
author Coelho, Beatriz Jorge
author_facet Coelho, Beatriz Jorge
Veigas, Bruno
Bettencourt, Luís
Águas, Hugo
Fortunato, Elvira
Martins, Rodrigo
Baptista, Pedro V.
Igreja, Rui
author_role author
author2 Veigas, Bruno
Bettencourt, Luís
Águas, Hugo
Fortunato, Elvira
Martins, Rodrigo
Baptista, Pedro V.
Igreja, Rui
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv DCM - Departamento de Ciência dos Materiais
UNINOVA-Instituto de Desenvolvimento de Novas Tecnologias
DCV - Departamento de Ciências da Vida
UCIBIO - Applied Molecular Biosciences Unit
RUN
dc.contributor.author.fl_str_mv Coelho, Beatriz Jorge
Veigas, Bruno
Bettencourt, Luís
Águas, Hugo
Fortunato, Elvira
Martins, Rodrigo
Baptista, Pedro V.
Igreja, Rui
dc.subject.por.fl_str_mv cancer biomarker
digital microfluidics
fluorescence detection
loop-mediated isothermal amplification
real-time nucleic acid amplification monitoring
Clinical Biochemistry
SDG 3 - Good Health and Well-being
topic cancer biomarker
digital microfluidics
fluorescence detection
loop-mediated isothermal amplification
real-time nucleic acid amplification monitoring
Clinical Biochemistry
SDG 3 - Good Health and Well-being
description LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy?i4HB. Additional funding was received from FCT under the scope of the project reference LA/P/0037/2020, Beatriz J. Coelho and grant COVID/BD/152453/2022. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.
publishDate 2022
dc.date.none.fl_str_mv 2022-08-03T22:26:02Z
2022-03-28
2022-03-28T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10362/142839
url http://hdl.handle.net/10362/142839
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 2079-6374
PURE: 43542197
https://doi.org/10.3390/bios12040201
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 12
application/pdf
dc.source.none.fl_str_mv reponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
instacron:RCAAP
instname_str FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
instacron_str RCAAP
institution RCAAP
reponame_str Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
collection Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
repository.name.fl_str_mv Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
repository.mail.fl_str_mv info@rcaap.pt
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