Cell-free DNA in maternal plasma and serum: A comparison of quantity, quality and tissue origin using genomic and epigenomic approaches
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摘要Objectives: The objectives of this study were to compare the concentrations, size profiles and major tissue contributors of cell-free DNA (cfDNA) in plasma and in serum.Design and methods: Thirteen pregnant women in the third trimester were recruited for this study. We collected EDTA-plasma and serum samples using various collection tubes. We determined their cfDNA concentrations and fetal cfDNA fractions using a zinc-finger X (ZFX)/zinc-finger Y (ZFY) droplet digital polymerase chain reaction (ZFX/ZFY ddPCR) assay. We used paired-end massively parallel sequencing (MPS) to measure plasma and serum cfDNA sizes at single-base resolution. We deconvoluted the genome-wide bisulfite sequencing data with reference to the methylation profiles of different tissues.Results: The concentrations of cfDNA collected in Sarstedt Serum Z tubes were found to be significantly higher than those in Greiner Bio-One Vacuette (R) Z Serum Separator Clot Activator tubes or Vacuette (R) Z Serum Clot Activator tubes. The concentrations of fetal cfDNA were significantly reduced in samples collected in the Vacuette serum collection tubes. Fetal cfDNA fractions were significantly reduced in all sera compared to plasma. MPS of serum cfDNA revealed a right shift of the size distributions compared to plasma. Methylation-based tissue mapping of serum cfDNA revealed an increase of cfDNA from neutrophils and B cells but not T cells.Conclusions: The use of different serum collection tubes has a significant impact on serum cfDNA concentrations. This effect is likely mediated through the combined effect of genomic DNA release from white blood cells and DNA degradation or removal. (C) 2016 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.
著者Wong FCK, Sun K, Jiang PY, Cheng YKY, Chan KCA, Leung TY, Chiu RWK, Lo YMD
期刊名稱Clinical Biochemistry
出版年份2016
月份12
卷號49
期次18
出版社PERGAMON-ELSEVIER SCIENCE LTD
頁次1379 - 1386
國際標準期刊號0009-9120
語言英式英語
關鍵詞Noninvasive prenatal testing, Serum cell-free DNA, Plasma cell-free DNA, Epigenetics, Digital PCR, Massively parallel sequencing, Pregnancy
Web of Science 學科類別Medical Laboratory Technology;Medical Laboratory Technology

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