胎儿游离DNA
真空吸尘器
DNA
数字聚合酶链反应
细胞
胎儿
聚合酶链反应
男科
分子生物学
实时聚合酶链反应
全血
产前诊断
化学
生物
怀孕
医学
遗传学
免疫学
色谱法
基因
作者
M. Rohan Fernando,K. Chen,Sharon A Norton,Gary Krzyzanowski,David W. A. Bourne,Bradford A. Hunsley,Wayne L. Ryan,Charlton H. Bassett
摘要
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 K(3)EDTA (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 K(3)EDTA 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.
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