title: Circulating cell free DNA: Preanalytical considerations.
abstract: Despite the growing interest in circulating cell-free DNA (ccfDNA) analysis in various clinical fields, especially oncology and prenatal diagnosis, few studies on sample handling have been reported and no analytical consensus is available. The lack of consistency between the various protocols for sample handling and the techniques used for ccfDNA analysis is one of the major obstacles in translating ccfDNA analysis to clinical practice. Although this point is highlighted regularly in the published reviews on ccfDNA analysis, no standard operating procedure currently exists despite several ongoing clinical studies on ccfDNA analysis. This review examines the preanalytical parameters potentially affecting ccfDNA concentration and fragmentation at each preanalytical step from blood drawing to the storage of ccfDNA extracts. Analysis of data in the literature and our own observations revealed the influence of preanalytical factors on ccfDNA analysis. Based on these data, we determined the optimal preanalytical protocols for ccfDNA analysis and ultimately, a guideline for the translation of ccfDNA analysis in routine clinical practice.
authors.affiliation: IRCM, Institut de Recherche en Cancérologie de Montpellier, Montpellier, F-34298, France; INSERM, U896, Montpellier, F-34298, France; Université Montpellier1, Montpellier, F-34298, France; Institut régional du Cancer Montpellier, Montpellier, F-34298, France.
authors.forename: Safia
authors.initials: S
authors.lastname: El Messaoudi
authors.forename: Fanny
authors.initials: F
authors.lastname: Rolet
authors.forename: Florent
authors.initials: F
authors.lastname: Mouliere
authors.forename: Alain R
authors.initials: AR
authors.lastname: Thierry
mesh_terms: D015415:Biomarkers / Q000097:blood; D001800:Blood Specimen Collection / Q000379:methods* / Q000592:standards; D004247:DNA / Q000097:blood*; D005615:Freezing; D006801:Humans; D012034:Refrigeration; D013997:Time Factors
chemical_list: D015415:Biomarkers; D004247:DNA
keywords: BRAF; Circulating cell-free DNA; DII; DNA integrity index; Diagnosis; EDTA; KRAS; Preanalytical handling; Q-PCR; RT; Stability; Standard operating procedure; ccfDNA; circulating cell-free DNA; ethylenediaminetetraacetic acid; quantitative polymerase chain reaction; room temperature; v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog [GenBank: NG_007524.1]; v-raf murine sarcoma viral oncogene homolog B1 [GenBank: NG_007873.2]