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Microfluidic extraction and digital quantification of circulating cell-free DNA from serum

Authors :
Iago Pereiro
Charlotte Proudhon
Sonia Garrigou
Karla Perez-Toralla
Jean-Louis Viovy
Valérie Taly
Stéphanie Descroix
François-Clément Bidard
Fahima Di Federico
Médecine Personnalisée, Pharmacogénomique, Optimisation Thérapeutique (MEPPOT - U1147)
Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)
Laboratoire Physico-Chimie Curie [Institut Curie] (PCC)
Institut Curie [Paris]-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)
Sorbonne Université (SU)
Laboratoire des biomarqueurs tumoraux circulants [Institut Curie, Paris] (SiRIC - Département de recherche translationnelle)
Institut Curie [Paris]-Université Paris sciences et lettres (PSL)
Institut Curie [Paris]
Physico-Chimie-Curie (PCC)
Institut Curie-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)
Université de recherche Paris-Sciences-et-Lettres (Université de recherche PSL)-Institut Curie [Paris]
Institut Curie
Source :
Sensors and Actuators B: Chemical, Sensors and Actuators B: Chemical, Elsevier, 2019, 286, pp.533-539. ⟨10.1016/j.snb.2019.01.159⟩
Publication Year :
2019
Publisher :
Elsevier BV, 2019.

Abstract

International audience; 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.

Details

ISSN :
09254005
Volume :
286
Database :
OpenAIRE
Journal :
Sensors and Actuators B: Chemical
Accession number :
edsair.doi.dedup.....da170f7e6e3ea2f9ef23dcb95cb8dacb