Journal
SENSORS AND ACTUATORS B-CHEMICAL
Volume 286, Issue -, Pages 533-539Publisher
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2019.01.159
Keywords
Microfluidic fluidized bed; Solid-phase extraction; Circulating cell-free DNA; Droplet digital PCR; Biomarkers
Funding
- SIRIC CARPEM
- Ministere de l'Enseignement Superieur et de la Recherche
- Universite Paris-Descartes
- Centre National de la Recherche Scientifique (CNRS)
- Institut National de la Sante et de la Recherche Medicale (INSERM)
- ANR Investissements d'Avenir for Digidiag project (ANR Nanobiotechnologies) [ANR-10-NANO-0002-09]
- ligue nationale contre le cancer (LNCC, Program Equipe labelisee LIGUE) [EL2016. LNCC/VaT]
- Labex
- Equipex IPGG
- CATCH-U-DNA
- FETOPEN H2020
- ERC Advanced Grant CellO [FP7-IDEAS-ERC-321107]
- European FP7 programs [LOVEFOOD FP7-ICT- 2011-317742, NAPES FP7-NMP-2013-604241]
- French Ile-de-France canceropole
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Miniaturized devices for the extraction of DNA have been used for assessing genetic material in biological, forensic and environmental samples. However, the ability to isolate trace amounts of highly fragmented DNA from biological fluids remains a challenge. The current work reports a microfluidic approach that combines on line a dynamic magnetic extraction procedure with droplet-based digital PCR (ddPCR). This strategy maximizes the surface area for DNA binding within the chip, in order to capture short DNA fragments, with the possibility of recovering the purified samples into picoliter volumes for high sensitivity mutation detection. The application of this technology to capture circulating cell-free DNA (ccfDNA) from serum samples of cancer patients is demonstrated herein, with efficiencies comparable to standard column-based DNA extraction methods. This technology uses lesser amounts of required material and reagents, and has a higher potential for automation and multiplex DNA analysis. Furthermore, this approach can also be extended for the detection of other circulating biomarkers, such as nucleic acid sequences with aberrant methylation patterns or miRNA.
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