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

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

Research output: Contribution to journalArticle

57 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 Rohan; 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 Rohan ; 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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