A new methodology to preserve the original proportion and integrity of cell-free fetal DNA in maternal plasma during sample processing and storage

M. R. Fernando, K. Chen, S. Norton, G. Krzyzanowski, D. Bourne, B. Hunsley, W. L. Ryan, C. Bassett

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

Objective: To develop a standardized blood collection device that preserves fetal cell-free DNA and minimizes the cell-free DNA background in maternal plasma. Methods: Blood samples were drawn from healthy pregnant donors into K3EDTA (BD vacutainer®) and Cell-free DNA™ BCT tubes (Streck®, Inc.) and kept at ambient temperature. Plasma was separated by centrifugation and cell-free DNA was extracted. Cell-free DNA from plasma was quantified by quantitative real-time polymerase chain reaction. Results: Blood drawn into Cell-free DNA™ BCT tubes showed no change in the original proportion of fetal cell-free DNA during a 14-day storage period at ambient temperature. Conversely, maternal blood drawn into K3EDTA tubes showed a steady reduction in the original proportion of fetal cell-free DNA over the same time period. Using maternal plasma stored in Cell-free DNA™ BCT tubes for 14 days, fetal cell-free DNA was amplified 80-fold using whole genome amplification (WGA). Conclusion: Using Streck's Cell-free DNA™ BCT tubes, it is possible to preserve the original proportion of fetal cell-free DNA for extended times as well as minimize the post-sampling maternal cell-free DNA background. Preserved in this way, fetal cell-free DNA can be amplified by WGA technology to be used in prenatal diagnostic tests.

Original languageEnglish (US)
Pages (from-to)418-424
Number of pages7
JournalPrenatal Diagnosis
Volume30
Issue number5
DOIs
StatePublished - May 1 2010

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Mothers
DNA
Genome
Temperature
Centrifugation
Routine Diagnostic Tests
Real-Time Polymerase Chain Reaction
Technology
Equipment and Supplies

Keywords

  • Cell-free
  • DNA
  • Fetal
  • Maternal
  • Plasma
  • Preservation

ASJC Scopus subject areas

  • Obstetrics and Gynecology
  • Genetics(clinical)

Cite this

A new methodology to preserve the original proportion and integrity of cell-free fetal DNA in maternal plasma during sample processing and storage. / Fernando, M. R.; Chen, K.; Norton, S.; Krzyzanowski, G.; Bourne, D.; Hunsley, B.; Ryan, W. L.; Bassett, C.

In: Prenatal Diagnosis, Vol. 30, No. 5, 01.05.2010, p. 418-424.

Research output: Contribution to journalArticle

Fernando, MR, Chen, K, Norton, S, Krzyzanowski, G, Bourne, D, Hunsley, B, Ryan, WL & Bassett, C 2010, 'A new methodology to preserve the original proportion and integrity of cell-free fetal DNA in maternal plasma during sample processing and storage', Prenatal Diagnosis, vol. 30, no. 5, pp. 418-424. https://doi.org/10.1002/pd.2484
Fernando, M. R. ; Chen, K. ; Norton, S. ; Krzyzanowski, G. ; Bourne, D. ; Hunsley, B. ; Ryan, W. L. ; Bassett, C. / A new methodology to preserve the original proportion and integrity of cell-free fetal DNA in maternal plasma during sample processing and storage. In: Prenatal Diagnosis. 2010 ; Vol. 30, No. 5. pp. 418-424.
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