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Tytuł:
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Single-use microfluidic device for purification and concentration of environmental DNA from river water.
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Autorzy:
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Carvalho J; International Iberian Nanotechnology Laboratory (INL), Avenida Mestre José Veiga, 4715-310, Braga, Portugal; Department of Analytical Chemistry, Nutrition and Food Science, College of Pharmacy/School of Veterinary Sciences, University of Santiago de Compostela, Campus Vida, E-15782, Santiago de Compostela, Spain.
Diéguez L; International Iberian Nanotechnology Laboratory (INL), Avenida Mestre José Veiga, 4715-310, Braga, Portugal.
Ipatov A; International Iberian Nanotechnology Laboratory (INL), Avenida Mestre José Veiga, 4715-310, Braga, Portugal.
Guerreiro JR; International Iberian Nanotechnology Laboratory (INL), Avenida Mestre José Veiga, 4715-310, Braga, Portugal.
Garrido-Maestu A; International Iberian Nanotechnology Laboratory (INL), Avenida Mestre José Veiga, 4715-310, Braga, Portugal.
Azinheiro S; International Iberian Nanotechnology Laboratory (INL), Avenida Mestre José Veiga, 4715-310, Braga, Portugal; Department of Analytical Chemistry, Nutrition and Food Science, College of Pharmacy/School of Veterinary Sciences, University of Santiago de Compostela, Campus Vida, E-15782, Santiago de Compostela, Spain.
Prado M; International Iberian Nanotechnology Laboratory (INL), Avenida Mestre José Veiga, 4715-310, Braga, Portugal. Electronic address: .
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Źródło:
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Talanta [Talanta] 2021 May 01; Vol. 226, pp. 122109. Date of Electronic Publication: 2021 Jan 21.
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Typ publikacji:
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Journal Article
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Język:
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English
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Imprint Name(s):
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Publication: Amsterdam : Elsevier
Original Publication: Oxford : Pergamon Press
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MeSH Terms:
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DNA, Environmental*
Microfluidic Analytical Techniques*
DNA/genetics ; Fresh Water ; Lab-On-A-Chip Devices ; Water
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Contributed Indexing:
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Keywords: DNA purification; Dreissena polymorpha; Invasive species; Microfluidic device; eDNA
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Substance Nomenclature:
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0 (DNA, Environmental)
059QF0KO0R (Water)
9007-49-2 (DNA)
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Entry Date(s):
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Date Created: 20210307 Date Completed: 20210514 Latest Revision: 20211204
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Update Code:
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20240104
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DOI:
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10.1016/j.talanta.2021.122109
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PMID:
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33676665
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Purification and concentration of DNA is a critical step on DNA-based analysis, which should ensure efficient DNA isolation and effective removal of contaminants that may interfere with downstream DNA amplification. Complexity of samples, minute content of target analyte, or high DNA fragmentation greatly entangles the success of this step. To overcome this issue, we designed and fabricated a novel miniaturized disposable device for a highly efficient DNA purification. The microfluidic device showed binding efficiency and elution yield of 90.1% and 86.7%, respectively. Moreover, the effect of DNA fragmentation, a parameter that has not been previously addressed, showed a great impact in the recovery step. The microfluidic system integrated micropillars with chitosan being used as the solid-phase for a pH-dependent DNA capture and release. We have showed the potential of the device in the successful purification of environmental DNA (eDNA) from river water samples contaminated with Dreissena polymorpha, an invasive alien species responsible for unquestionable economic and environmental consequences in river water basins. Additionally, the device was also able to concentrate the DNA extract from highly diluted samples, showing promising results for the early detection of such invasive species, which may allow prompt measures for a more efficient control in affected areas. Suitability for integration with downstream DNA analysis was also demonstrated through qPCR analysis of the samples purified with the microfluidic device, allowing detection of the target species even if highly diluted.
(Copyright © 2021 Elsevier B.V. All rights reserved.)