Journal article
Effects of Collection and Processing Procedures on Plasma Circulating Cell-Free DNA from Cancer Patients
B Risberg, DWY Tsui, H Biggs, A Ruiz-Valdepenas Martin de Almagro, SJ Dawson, C Hodgkin, L Jones, C Parkinson, A Piskorz, F Marass, D Chandrananda, E Moore, J Morris, V Plagnol, N Rosenfeld, C Caldas, JD Brenton, D Gale
Journal of Molecular Diagnostics | ELSEVIER SCIENCE INC | Published : 2018
Open access
Abstract
Circulating tumor DNA (ctDNA) offers new opportunities for noninvasive cancer management. Detecting ctDNA in plasma is challenging because it constitutes only a minor fraction of the total cell-free DNA (cfDNA). Pre-analytical factors affect cfDNA levels contributed from leukocyte lysis, hence the ability to detect low-frequency mutant alleles. This study investigates the effects of the delay in processing, storage temperatures, different blood collection tubes, centrifugation protocols, and sample shipment on cfDNA levels. Peripheral blood (n = 231) from cancer patients (n = 62) were collected into K3EDTA or Cell-free DNA BCT tubes and analyzed by digital PCR, targeted amplicon, or shallow ..
View full abstractGrants
Awarded by National Breast Cancer Foundation
Funding Acknowledgements
We thank the patients for consenting to participate, the Human Research Tissue Bank at Addenbrooke's Hospital, which is supported by the NIHR Cambridge Biomedical Research Center, Frank Diehl for discussions about method standardization, Irena Hudecova for help preparing figures, and the Genomics Core of the Cancer Research UK Cambridge Institute for providing sequencing support.