title: Comprehensive evaluation of methods to isolate, quantify, and characterize circulating cell-free DNA from small volumes of plasma.
abstract: Circulating cell-free DNA (ccfDNA) has great potential for non-invasive diagnostics, and prediction and monitoring of treatment response, but its amount is usually limited. Therefore, the choice of methods to extract and characterize ccfDNA is crucial. In the current study, we performed the most comprehensive comparison of methods for ccfDNA extraction (11 methods), quantification (3 methods), and estimation of the integrity index (2 methods) from small quantities of different kinds of plasma. The QIAamp® Circulating Nucleic Acid Kit and the Norgen Plasma/Serum Circulating DNA Purification Mini Kit showed the best accuracy and reproducibility, but the Norgen kit allowed to extract a higher amount of ccfDNA. This workflow provides a reliable protocol for the multiple applications of ccfDNA in biomedicine.
authors.affiliation: Laboratory for Epigenetics and Environment, Centre National de Génotypage, CEA-Institut de Génomique, 2 rue Gaston Crémieux, CP 5721, 91000, Evry, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Mauger
authors.forename: Cécile
authors.initials: C
authors.lastname: Dulary
authors.forename: Christian
authors.initials: C
authors.lastname: Daviaud
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Deleuze
authors.forename: Jorg
authors.initials: J
authors.lastname: Tost
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D002474:Cell-Free System; D004273:DNA, Neoplasm / Q000097:blood* / Q000235:genetics* / Q000302:isolation & purification; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D004273:DNA, Neoplasm