NIH, National Cancer Institute, Division of Cancer Treatment and Diagnosis (DCTD) NIH - National Institutes of Health National Cancer Institute DCTD - Division of Cancer Treatment and Diagnosis

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

The purpose of this paper was to determine the effects of duration at room temperature, storage temperature, mechanical stress, and collection tube type on the stability of cell free DNA in plasma from pregnant women.

Conclusion of Paper

Total and fetal circulating cell free DNA concentrations in plasma were unchanged when blood was stored for up to 7 days at room temperature in Streck tubes which contain EDTA, but total circulating cell-free DNA in plasma increased when blood was stored at room temperature for 2 days or more in EDTA tubes. While fetal circulating cell free DNA concentrations in plasma were unaffected by storage at any temperature examined, total circulating cell free DNA in plasma increased when whole blood was stored at 23 degrees C or higher. There was no effect of dropping the box containing whole blood in Streck tubes or tube lot number on levels of total or fetal circulating cell free DNA in plasma.

Studies

  1. 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:
    AnalyteTechnology Platform
    DNA MALDI-TOF MS
    DNA DNA sequencing
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(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
  2. 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:
    AnalyteTechnology Platform
    DNA MALDI-TOF MS
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(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

You Recently Viewed  

comments powered by Disqus

News and Announcements

  • Most Popular SOPs in 2018

  • New Article Published from NCI's Biospecimen Pre-analytical Variables (BPV) Program

  • 2019 Annual ISBER Meeting

  • More...