Optimizing blood collection, transport and storage conditions for cell free DNA increases access to prenatal testing.
Author(s): Wong D, Moturi S, Angkachatchai V, Mueller R, Desantis G, van den Boom D, Ehrich M
Publication: Clin Biochem, 2013, Vol. 46, Page 1099-104
PubMed ID: 23643886 PubMed Review Paper? No
Purpose of Paper
Conclusion of Paper
Studies
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Study Purpose
The purpose of this study was to determine the effects of up to 14 days of room temperature storage of whole blood in EDTA or Streck tubes on the levels of circulating cell-free total and fetal DNA in plasma from pregnant women. Blood collected in EDTA tubes was transported on ice packs, and plasma was either separated within 6 h or after 1-14 days. Streck tubes are a proprietary tube containing EDTA. Blood collected in Streck tubes was shipped at ambient temperatures and plasma was either separated within 24 h or after 1-14 days. All plasma was frozen before analysis. DNA was extracted from frozen plasma using QIAamp DSP circulating NA kit
Summary of Findings:
Total and fetal circulating cell-free DNA remained constant during the first 7 and 14 days of room temperature storage of blood in Streck tubes, respectively, but total circulating cell-free DNA increased slightly by day 14 compared to day 0 (p<0.0001). In contrast, whole blood that was collected and stored in EDTA tubes had increased total circulating cell-free DNA by day 2 of storage (p=0.0007) and increased fetal circulating cell-free DNA by day 14 of storage (p=0.0050). Further, while the ratio of fetal to total circulating cell-free DNA was stable until day 7 in specimens stored in Streck tubes, it was significantly lower between days 2 and 7 in the EDTA specimens (p<0.05, all). Streck tube lot number had no effect on levels of total and fetal circulating cell-free DNA. Further, there were no differences in DNA concentrations of sequencing libraries produced from blood collected in Streck and EDTA tubes.
Biospecimens
Preservative Types
- Streck/BCT
- None (Fresh)
Diagnoses:
- Pregnant
Platform:
Analyte Technology Platform DNA MALDI-TOF MS DNA DNA sequencing Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Time at room temperature 0 days
1 day
2 days
3 days
4 days
7 days
14 days
Storage Between site transportation method Unspecified transport method
Mailed
Biospecimen Acquisition Type of collection container/solution EDTA tubes
Streck tubes
Multiple Streck lots compared
Biospecimen Aliquots and Components Centrifugation Centrifugation delays investigated
Biospecimen Preservation Type of fixation/preservation Refrigeration
Blood collection tube additive
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Study Purpose
The purpose of this study was to determine the effects of storage temperature and dropping whole blood in Streck tubes on the levels of circulating cell-free total and fetal DNA in plasma from pregnant women. Streck tubes are a proprietary tube containing EDTA. Blood collected in Streck tubes was shipped at ambient temperatures, and plasma was isolated within 30 h (0 h) or after a further 24 h at the experimental temperature. To investigate the effects of handling, specimens were packaged according to the International Air Transport Association (IATA) regulations for transport of blood specimens and then dropped 3 feet to the floor. All plasma was frozen before analysis. DNA was extracted from frozen plasma using QIAamp DSP circulating NA kit.
Summary of Findings:
Copies of total circulating cell-free DNA increased when specimens were stored for an additional 24 h at 23, 37 or 40 degrees C compared to specimens that were not subjected to storage after arrival at the laboratory (p<0.001, all), but copies of fetal circulating cell-free DNA were not affected by further storage regardless of temperature. Consequently, the fetal fraction of circulating cell-free DNA was lower in specimens stored at 23, 37 or 40 degrees C for 24 h than specimens that were not subjected to additional storage (p<0.001, all). There was no effect of dropping the shipping box containing blood from 3 feet on the total or fetal circulating cell-free DNA content.
Biospecimens
Preservative Types
- Streck/BCT
- None (Fresh)
Diagnoses:
- Pregnant
Platform:
Analyte Technology Platform DNA MALDI-TOF MS Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage temperature 4 degrees C
23 degrees C
37 degrees C
40 degrees C
Storage Storage conditions Dropped from 3 feet
Not dropped
Biospecimen Aliquots and Components Centrifugation Centrifugation delays investigated