A stabilizing reagent prevents cell-free DNA contamination by cellular DNA in plasma during blood sample storage and shipping as determined by digital PCR.

Author(s): Norton SE, Lechner JM, Williams T, Fernando MR

Publication: Clin Biochem, 2013, Vol. 46, Page 1561-5

PubMed ID: 23769817 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of collection, storage and transport of whole blood in Streck BCT tubes (BCT) rather than K3 EDTA tubes on levels of cell-free and genomic DNA in plasma.

Conclusion of Paper

When whole blood was stored in K3 EDTA tubes at room temperature, significant increases in cell-free and genomic DNA were noted in plasma on day 2 and day 1, respectively. However, when whole blood was stored at room temperature in BCT tubes, plasma levels of cell-free and genomic DNA were stable for 7 days, but they were higher after 14 days than on day 0. When whole blood was shipped round-trip by air in K3EDTA tubes, cell-free and genomic DNA levels in plasma were higher than prior to shipment (p=0.0011 and 0.0157, respectively), but levels of cell-free and genomic DNA in plasma were not affected by shipment in BCT tubes.

Studies

Study Purpose

The purpose of this study was to determine the effects of collection, storage and transport of whole blood in BCT rather than K3 EDTA tubes on levels of cell-free and genomic DNA in plasma. Research was conducted at the manufacturer of the BCT tubes. Blood was shipped round-trip in insulated boxes by overnight air freight between Omaha and Lincoln, Nebraska. DNA was extracted with QIAamp Circulating Nucleic Acid Kit and stored at -60 degrees C until analysis. DNA was quantified by digital PCR amplification of a 136 bp (cell-free DNA) and 420 bp (genomic DNA) fragments of beta actin.

Summary of Findings:

When whole blood was stored in K3 EDTA tubes at room temperature, significant increases in cell-free and genomic DNA were noted in plasma on day 2 (2 fold, p=0.0092) and day 1 (1.2 fold, p=0.037), respectively, compared to day 0. However, when whole blood was stored at room temperature in BCT tubes, plasma levels of cell-free and genomic DNA were stable for 7 days, but after 14 days, they were 4 (p<0.0001) and 1.9 fold (p<0.02) higher, respectively, than on day 0. When whole blood was shipped round-trip by air in K3EDTA tubes, cell-free and genomic DNA levels in plasma were higher than prior to shipment (p=0.0011 and 0.0157, respectively), but levels of cell-free and genomic DNA were not affected by shipment in BCT tubes.

Biospecimens

Preservative Types

None (Fresh)
Streck/BCT

Diagnoses:

Normal

Platform:

Analyte	Technology Platform
DNA	Digital PCR

Pre-analytical Factors:

Classification	Pre-analytical Factor	Value(s)
Biospecimen Acquisition	Type of collection container/solution	K3EDTA vacutainer Streck BCT
Storage	Between site transportation method	Air courier Not transported
Storage	Time at room temperature	0 days 1 day 2 days 3 days 7 days 14 days
Biospecimen Aliquots and Components	Blood and blood products	Plasma Whole blood
Digital PCR Specific	Targeted nucleic acid	Beta-actin
Biospecimen Aliquots and Components	Centrifugation	Centrifugation delays investigated
Biospecimen Preservation	Type of fixation/preservation	Blood collection tube additive None (fresh)

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