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

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 ke...

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Bibliographic Details
Published in:Prenatal diagnosis 2010-05, Vol.30 (5), p.418-424
Main Authors: Fernando, M. R., Chen, K., Norton, S., Krzyzanowski, G., Bourne, D., Hunsley, B., Ryan, W. L., Bassett, C.
Format: Article
Language:English
Subjects:
Biological and medical sciences
Blood Specimen Collection - methods
Blood Specimen Collection - standards
Case-Control Studies
cell-free
Centrifugation
Chromosomes, Human, Y - genetics
Delivery. Postpartum. Lactation
DNA
DNA - blood
Female
fetal
Fetuses
Fundamental and applied biological sciences. Psychology
Genetics of eukaryotes. Biological and molecular evolution
Genomes
Gynecology. Andrology. Obstetrics
Humans
maternal
Maternal-Fetal Exchange - genetics
Medical sciences
Molecular and cellular biology
plasma
Polymerase Chain Reaction
Pregnancy
Pregnancy Trimester, First - genetics
Prenatal diagnosis
preservation
Temperature effects
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Summary: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. Copyright © 2010 John Wiley & Sons, Ltd.
ISSN:0197-3851
1097-0223
1097-0223
DOI:10.1002/pd.2484