A new blood collection device minimizes cellular DNA release during sample storage and shipping when compared to a standard device.
Author(s): Norton SE, Luna KK, Lechner JM, Qin J, Fernando MR
Publication: J Clin Lab Anal, 2013, Vol. 27, Page 305-11
PubMed ID: 23852790 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to determine the effects of the blood collection tube type or use of Streck blood collection tube (BCT) stabilizing reagent, delayed centrifugation, shipment, plasma storage, and genomic DNA concentration on cell-free DNA levels in plasma.
Conclusion of Paper
Significant increases in plasma cell-free DNA concentrations were observed when blood was stored in K3EDTA tubes for 14 days at 22°C, 7 or 14 days at 37°C, or on an orbital shaker for 6 h at 22°C before centrifugation, but the same treatments had no effect when blood was stored in Streck BCT tubes. Further, cell-free DNA concentrations in plasma were increased when blood specimens were shipped in K3EDTA tubes prior to centrifugation, but not Streck BCT tubes. Detection of exogenous SRY was inhibited when blood was stored at 22°C in K3EDTA tubes but not in BCT tubes. Storage of plasma at room temperature with the BCT additive for 7 or 14 days affected levels of cell-free DNA.
Studies
-
Study Purpose
The purpose of this study was to determine the effect of adding the BCT stabilizing reagent to plasma on levels of cell-free DNA after room temperature storage. Blood from 5 donors was collected into EDTA tubes. Plasma was obtained by double centrifugation and pooled into new tubes. Pooled plasma was stored with or without the BCT additive at 22° for 3 h, 6 h, 7 days or 14 days. DNA was isolated using the QIAamp Circulating Nucleic Acids kit. Plasma and isolated DNA were stored at -80°C.
Summary of Findings:
Addition of the BCT cell stabilizing agent to plasma had no effect on cell-free DNA levels in plasma stored at 22°C for 3 h, or 6 h, but after 7 days, plasma with the BCT additive had lower levels of cell-free DNA than that stored without (p=0.0002), and after 14 days, plasma with the BCT additive had slightly higher levels of cell-free DNA than that stored without (p=0.029).
Biospecimens
Preservative Types
- Streck/BCT
- None (Fresh)
Diagnoses:
- Normal
Platform:
Analyte Technology Platform DNA Real-time qPCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Time at room temperature 3 h
6 h
7 days
14 days
Real-time qPCR Specific Targeted nucleic acid Beta-actin
Biospecimen Preservation Type of fixation/preservation Blood collection tube additive
None (fresh)
-
Study Purpose
The purpose of this study was to determine the effects of increasing genomic DNA, as occurs with delayed plasma separation, on the detection of exogenous DNA in plasma from BCT and K3EDTA tubes. Blood from 5 healthy women was stored at room temperature for 0, 7 or 14 days. Plasma was obtained by double centrifugation and stored frozen at -80°C. Thawed plasma was spiked with plasmid DNA containing the SRY gene, and DNA was isolated using the QIAamp Circulating Nucleic Acids kit.
Summary of Findings:
When blood was stored in K3EDTA or Streck BCT tubes for 14 days rather than 0 days prior to centrifugation and separation of plasma, 92-fold and 8-fold increases in beta-actin copies, respectively, were noted (p=0.0091 and p=0.0090, respectively) in DNA isolated from plasma. Corresponding decreases in SRY detection were found in specimens stored for 7 days (3-fold) or 14 days (95-fold) in K3EDTA tubes (p=0.0055 and p=0.0006, respectively), but SRY detection in plasma was not altered when blood was stored in Streck BCT tubes.
Biospecimens
Preservative Types
- Streck/BCT
- None (Fresh)
Diagnoses:
- Normal
Platform:
Analyte Technology Platform DNA Real-time qPCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Time at room temperature 0 days
7 days
14 days
Biospecimen Aliquots and Components Centrifugation Centrifugation delays investigated
Biospecimen Acquisition Type of collection container/solution Streck BCT
K3EDTA tube
Real-time qPCR Specific Targeted nucleic acid Beta-actin
SRY
Biospecimen Preservation Type of fixation/preservation Blood collection tube additive
None (fresh)
-
Study Purpose
The purpose of this study was to determine the effects of collection tube type, and shaking, room temperature storage and transport of blood on levels of cell-free DNA in plasma. For the shaking experiment, blood from 8 donors was put on an orbital shaker at 150 rpm and centrifuged at 0, 3, 6, or 24 h. To study the effects of shipment, blood from 10 donors was: i) centrifuged within 2 h, ii) shipped in an insulated cooler during winter from Nebraska to Massachusetts and back (4 days), or iii) stored 4 days at 22°C before centrifugation. Plasma was obtained by double centrifugation, and DNA was isolated using the QIAamp Circulating Nucleic Acids kit. Plasma and isolated DNA were stored at -80°C.
Summary of Findings:
When blood was stored in K3EDTA tubes on an orbital shaker at 22°C, the levels of cell free DNA in plasma increased linearly and were significantly higher after 6 h than at 0 h (p=0.0015). Compared to baseline levels measured in the respective tubes, significantly higher levels of cell-free DNA were found in plasma from blood stored on an orbital shaker in a Streck BCT tube for 3 h (p=0.0182) or in non-shaken blood stored in a K3EDTA tube for 3 h (p=0.0029). However, cell-free DNA levels in plasma were not affected by 6-24 h storage of blood at 22°C in BCT tubes, with or without shaking or in K3EDTA tubes without shaking. While initial levels of cell-free DNA were comparable among plasma from BCT and K3EDTA tubes, plasma from blood shipped for 4 days in an insulated cooler or stored for 4 days at 22°C in K3EDTA tubes had higher levels of cell-free DNA compared to initial levels (p=0.0155 and p=0.028, respectively), and higher levels of cell-free DNA were found in plasma from blood shipped round-trip in K3EDTA rather than BCT tubes (p=0.0076).
Biospecimens
Preservative Types
- Streck/BCT
- None (Fresh)
Diagnoses:
- Normal
Platform:
Analyte Technology Platform DNA Real-time qPCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Type of collection container/solution K3EDTA tube
BCT
Biospecimen Aliquots and Components Centrifugation Centrifugation delays investigated
Storage Storage duration 0 h
3 h
6 h
24 h
0 days
4 days
Storage Between site transportation method Mailed
Not transported
Biospecimen Aliquots and Components Biospecimen mixing 150 rpm on orbital shaker
Not shaken
Real-time qPCR Specific Targeted nucleic acid Beta-actin
Storage Storage conditions 150 rpm on orbital shaker
Not shaken
Biospecimen Preservation Type of fixation/preservation Blood collection tube additive
None (fresh)
-
Study Purpose
The purpose of this study was to investigate the effects of blood storage duration and temperature on plasma cell-free DNA levels. Blood from 6 donors was stored for 0, 7 or 14 days at 6°C, 22°C, or 37°C. After storage, plasma was obtained by double centrifugation and stored at -80°C. DNA was isolated using the QIAamp Circulating Nucleic Acids kit and stored at -80°C.
Summary of Findings:
Large increases in plasma cell-free DNA concentrations were observed when blood was stored in K3EDTA tubes for 14 days at 6°C (78-fold), 22°C (233-fold), or 37°C (53-fold), but the increases were only significant when stored at 22°C (p=0.0028 at 14 days) or 37°C (30-fold, p=0.0104 at 7 days and p=0.009 at 14 days). In contrast, levels of cell-free DNA were not affected by up to 14 days of storage in Streck BCT tubes, regardless of temperature.
Biospecimens
Preservative Types
- Streck/BCT
- None (Fresh)
Diagnoses:
- Normal
Platform:
Analyte Technology Platform DNA Real-time qPCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Acquisition Type of collection container/solution BCT
K3EDTA tube
Biospecimen Aliquots and Components Centrifugation Centrifugation delays investigated
Storage Storage duration 0 days
7 days
14 days
Storage Storage temperature 6°C
22°C
37°C
Real-time qPCR Specific Targeted nucleic acid Beta-actin
Biospecimen Preservation Type of fixation/preservation Blood collection tube additive
None (fresh)