Effects of Collection and Processing Procedures on Plasma Circulating Cell-Free DNA from Cancer Patients.

摘要: 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 (cfDNA). Pre-analytical factors affect cfDNA levels contributed from leukocyte lysis, hence ability to detect low-frequency mutant alleles. This study investigates effects delay processing, storage temperatures, different blood collection tubes, centrifugation protocols, and sample shipment on levels. Peripheral (n = 231) patients (n = 62) were collected into K3EDTA or Cell-free BCT tubes analyzed by digital PCR, targeted amplicon, shallow whole-genome sequencing. To assess pre-analytic effects, was processed under conditions after 0, 6, 24, 48, 96 hours, 1 week at room temperature 4°C, using protocols. Digital PCR showed that increased gradually with time but stable tubes. samples stored 4°C less variation than storage, elevated compared BCT. A second 3000 × g gave similar yields higher-speed centrifugation. Next-generation sequencing negligible differences background error copy number changes between BCT, following provides insights sample processing analysis.