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BIABooster: Online DNA Concentration and Size Profiling with a Limit of Detection of 10 fg/μL and Application to High-Sensitivity Characterization of Circulating Cell-Free DNA

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posted on 02.03.2018, 00:00 by Comtet-Louis Andriamanampisoa, Aurélien Bancaud, Audrey Boutonnet-Rodat, Audrey Didelot, Jacques Fabre, Frédéric Fina, Fanny Garlan, Sonia Garrigou, Caroline Gaudy, Frédéric Ginot, Daniel Henaff, Pierre Laurent-Puig, Arnaud Morin, Vincent Picot, Laure Saias, Valérie Taly, Pascale Tomasini, Aziz Zaanan
We describe a technology to perform sizing and concentration analysis of double stranded DNA with a sensitivity of 10 fg/μL in an operating time of 20 min. The technology is operated automatically on a commercial capillary electrophoresis instrument using electro-hydrodynamic actuation. It relies on a new capillary device that achieves online concentration of DNA at the junction between two capillaries of different diameters, thanks to viscoelastic lift forces. Using a set of DNA ladders in the range of 100–1500 bp, we report a sizing accuracy and precision better than 3% and a concentration quantification precision of ∼20%. When the technology is applied to the analysis of clinical samples of circulating cell-free DNA (cfDNA), the measured cfDNA concentrations are in good correlation with those measured by digital PCR. Furthermore, the cfDNA size profiles indicate that the fraction of low molecular weight cfDNA in the range of 75–240 bp is a candidate biomarker to discriminate between healthy subjects and cancer patients. We conclude that our technology is efficient in analyzing highly diluted DNA samples and suggest that it will be helpful in translational and clinical research involving cfDNA.

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