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Valveless On-Chip Aliquoting for Molecular Diagnosis

The detection of nucleic acids as specific markers of infectious diseases is commonly implemented in molecular biology laboratories. The translation of these benchtop assays to a lab-on-a-chip format demands huge efforts of integration and automation. The present work is motivated by a strong requir...

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Autores principales: Romero Deza, Andersson A., Schaumburg, Federico, Berli, Claudio L. A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10386703/
https://www.ncbi.nlm.nih.gov/pubmed/37512736
http://dx.doi.org/10.3390/mi14071425
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author Romero Deza, Andersson A.
Schaumburg, Federico
Berli, Claudio L. A.
author_facet Romero Deza, Andersson A.
Schaumburg, Federico
Berli, Claudio L. A.
author_sort Romero Deza, Andersson A.
collection PubMed
description The detection of nucleic acids as specific markers of infectious diseases is commonly implemented in molecular biology laboratories. The translation of these benchtop assays to a lab-on-a-chip format demands huge efforts of integration and automation. The present work is motivated by a strong requirement often posed by molecular assays that combine isothermal amplification and CRISPR/Cas-based detection: after amplification, a 2–8 microliter aliquot of the reaction products must be taken for the subsequent reaction. In order to fulfill this technical problem, we have designed and prototyped a microfluidic device that is able to meter and aliquot in the required range during the stepped assay. The operation is achieved by integrating a porous material that retains the desired amount of liquid after removing the excess reaction products, an innovative solution that avoids valving and external actuation. The prototypes were calibrated and experimentally tested to demonstrate the overall performance (general fluidics, metering, aliquoting, mixing and reaction). The proposed aliquoting method is fully compatible with additional functions, such as sample concentration or reagent storage, and could be further employed in alternative applications beyond molecular diagnosis.
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spelling pubmed-103867032023-07-30 Valveless On-Chip Aliquoting for Molecular Diagnosis Romero Deza, Andersson A. Schaumburg, Federico Berli, Claudio L. A. Micromachines (Basel) Article The detection of nucleic acids as specific markers of infectious diseases is commonly implemented in molecular biology laboratories. The translation of these benchtop assays to a lab-on-a-chip format demands huge efforts of integration and automation. The present work is motivated by a strong requirement often posed by molecular assays that combine isothermal amplification and CRISPR/Cas-based detection: after amplification, a 2–8 microliter aliquot of the reaction products must be taken for the subsequent reaction. In order to fulfill this technical problem, we have designed and prototyped a microfluidic device that is able to meter and aliquot in the required range during the stepped assay. The operation is achieved by integrating a porous material that retains the desired amount of liquid after removing the excess reaction products, an innovative solution that avoids valving and external actuation. The prototypes were calibrated and experimentally tested to demonstrate the overall performance (general fluidics, metering, aliquoting, mixing and reaction). The proposed aliquoting method is fully compatible with additional functions, such as sample concentration or reagent storage, and could be further employed in alternative applications beyond molecular diagnosis. MDPI 2023-07-15 /pmc/articles/PMC10386703/ /pubmed/37512736 http://dx.doi.org/10.3390/mi14071425 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Romero Deza, Andersson A.
Schaumburg, Federico
Berli, Claudio L. A.
Valveless On-Chip Aliquoting for Molecular Diagnosis
title Valveless On-Chip Aliquoting for Molecular Diagnosis
title_full Valveless On-Chip Aliquoting for Molecular Diagnosis
title_fullStr Valveless On-Chip Aliquoting for Molecular Diagnosis
title_full_unstemmed Valveless On-Chip Aliquoting for Molecular Diagnosis
title_short Valveless On-Chip Aliquoting for Molecular Diagnosis
title_sort valveless on-chip aliquoting for molecular diagnosis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10386703/
https://www.ncbi.nlm.nih.gov/pubmed/37512736
http://dx.doi.org/10.3390/mi14071425
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