Concordance of Mutation Detection in Circulating Tumor DNA in Early Clinical Trials Using Different Blood Collection Protocols.
Author(s): Ahlborn LB, Madsen M, Jonson L, Nielsen FC, Lassen U, Yde CW, Mau-Sorensen M
Publication: Clin Lab, 2017, Vol. 63, Page 1755-1759
PubMed ID: 29035458 PubMed Review Paper? No
Purpose of Paper
This paper compared the fractional abundance of BRAF and PIK3CA mutations in plasma from blood collected in cell-free DNA BCT tubes and stored for up to 72 h before processing with that in matched specimens collected in EDTA tubes and processed within 15 min.
Conclusion of Paper
The fractional abundance of each mutation was comparable between tube types for each of the specimens.
Studies
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Study Purpose
This study compared the fractional abundance of BRAF and PIK3CA mutations in plasma from blood collected in cell-free DNA BCT tubes and stored for up to 72 h before processing with that in matched specimens collected in EDTA tubes and processed within 15 min. Blood was collected from 5 patients with solid tumors (colorectal, bile duct, lung, and breast cancers) at 4-6 different timepoints (before and during treatments) into cell-free DNA BCT and EDTA vacuette tubes. Plasma was obtained from specimens in EDTA tubes within 15 min by centrifugation at 1600 x g for 10 min followed by 20,000 x g for 20 min. Plasma was obtained from specimens in BCT tubes within 72 h by centrifugation at 2250 x g for 20 min followed by 18,000 x g for 20 min. Plasma was stored at -80˚C until DNA extraction using the QIAsymphony Circulating DNA kit. DNA was quantified using the dsDNA HS Qubit assay and stored at -20˚C. Mutant and wildtype copies were quantified using at least 10,000 drops in droplet digital PCR.
Summary of Findings:
The fractional abundance of each mutation was comparable between tube types for each of the specimens. Further, no systemic effect of collection tube type was observed.
Biospecimens
Preservative Types
- Frozen
- Streck/BCT
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform DNA Digital PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Type of collection container/solution Streck BCT
EDTA Vacuette
Biospecimen Preservation Type of fixation/preservation Blood collection tube additive
EDTA
Storage Time at room temperature <15 min
<72 h
Digital PCR Specific Targeted nucleic acid PIK3CA
BRAF