The In Vitro Stability of Circulating Tumour DNA.
Author(s): Henao Diaz E, Yachnin J, Grönberg H, Lindberg J
Publication: PLoS One, 2016, Vol. 11, Page e0168153
PubMed ID: 27959945 PubMed Review Paper? No
Purpose of Paper
This paper compared the stability of circulating tumor DNA (ctDNA) in blood collected in EDTA and Streck BCT tubes.
Conclusion of Paper
The ctDNA fraction and detection of copy number changes was unaffected by storage for 48 h after collection in either tube type. While significant differences in ONK5 amplification after storage at 5˚C for 48 h and in ONK15 deletion after storage at 5˚C for 24 h, these changes were not associated with any change in ctDNA signal and the proportion of each mutation was unchanged.
Studies
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Study Purpose
This study investigated the stability of ctDNA in blood collected in EDTA and Streck BCT tubes. Blood obtained from nine patients with metastatic prostate cancer in EDTA and Streck cell-free DNA BCT tubes was aliquoted into microcentrifuge tubes. Specimens from six patients were stored at room temperature (EDTA and Streck BCT tubes) or 5˚C (EDTA tubes only) for 0, 2, 3.2-6, and 24 h and specimens from three patients were either processed immediately or stored at room temperature for 48 h. Plasma was obtained by centrifugation at 1100 x g for 10 min followed by 1600 x g for 10 min and stored at -80˚C until DNA extraction using the QIAamp Circulating Nucleic Acid kit. Libraries were constructed using the ThruPLEX DNA-seq kit and sequenced using the EZ SeqCap kit on an Illumina Hiseq 2500.
Summary of Findings:
The ctDNA fraction and detection of copy number changes was unaffected by storage for 48 h iregardless of collection tube type. Compared to baseline, a significant difference was found in ONK5 amplification after storage at 5˚C for 48 h (P=0.02) and in ONK15 deletion after storage at 5˚C for 24 h (P=0.0003). However, these changes were not associated with any change in ctDNA signal and the proportion of each mutation was unchanged.
Biospecimens
Preservative Types
- None (Fresh)
- Streck/BCT
Diagnoses:
- Neoplastic - Carcinoma
Platform:
Analyte Technology Platform DNA Nucleoid sedimentation assay Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Type of collection container/solution EDTA tube
Streck BCT
Storage Storage temperature 5˚C
Room temperature
Storage Storage duration 0 h
2 h
3.2-6 h
24 h
48 h
Biospecimen Preservation Type of fixation/preservation EDTA
Blood collection tube additive