Detection of cell-free fetal DNA in maternal urine
Detection of cell-free fetal DNA in maternal urine
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DOI：
10.1002/pd.1591
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发表时间：
2006-12-01
影响因子：
3
通讯作者：
Finning, K.
中科院分区：
文献类型：
--
作者：
Illanes, S.;Denbow, M. L.;Finning, K.
目的探讨孕妇尿中无细胞胎儿DNA信号作为无创性产前诊断材料的潜在来源。母血和尿液以10000 g离心10分钟。血浆和尿液上清液各1m L，移至干净的试管中，再次离心。在不干扰细胞颗粒的情况下，去除血浆(0.8mL)和尿液(0.8mL)上清液，并将其保存在-80℃。提取DNA后，用实时定量聚合酶链式反应(PCR)检测Y染色体相关DYS14基因的存在。每个样本中的DNA总量(母体和胎儿)通过实时定量聚合酶链式反应(RT-PCR)进行估计。CVS的胎龄中位数为13周(范围11+5-14+1)。经核型分析证实，胎儿为男性12例，女性8例。在20个孕妇尿样中均未检测到Y染色体DNA，包括在母体血浆中检测到Y染色体DNA的20个样本C中的12个(随后均被证实怀有男性胎儿)。结论孕妇尿液中无细胞胎儿DNA存在或扩增不明显。版权所有(C)2006 John Wiley&Sons，Ltd.
Objective To evaluate the presence of cell-free fetal DNA signals in maternal urine as a potential source of material for non-invasive prenatal diagnosis.Study Design Patients referred to the regional fetal medicine unit who underwent prenatal diagnosis by chorionic villus sampling (CVS) were asked to give blood and urine immediately before the procedure. Maternal blood and urine were centrifuged at 10000 g for 10 min. Plasma (1 mL) and urine (1 mL) supernatant were transferred to a clean tube and centrifuged again. The plasma (0.8 mL) and urine (0.8 mL) supernatant were removed without disturbing the cell pellet and stored at -80 degrees C. Following DNA extraction, each sample was tested for the presence of Y chromosome associated DYS14 gene using real-time polymerase chain reaction (PCR). The total amount (maternal and fetal) of DNA in each sample was estimated using a quantitative real-time PCR assay.Results Twenty patients were enrolled in the study. CVS was performed at a median gestational age of 13 weeks (range 11 + 5-14 + 1). There were 12 male and 8 female fetuses, as confirmed by karyotype. Y chromosome DNA was not detected in any of the 20 samples of maternal urine, including 12 of the 20 samples C in which Y chromosome DNA was detected in maternal plasma (all of whom were subsequently confirmed to be carrying a male fetus). There was considerable variation in the amount of total free DNA detected in maternal urine.Conclusions Cell-free fetal DNA either was not present or did not amplify in maternal urine. Copyright (c) 2006 John Wiley & Sons, Ltd.