Accurate and robust quantification of circulating fetal and total DNA in maternal plasma from 5 to 41 weeks of gestation
Accurate and robust quantification of circulating fetal and total DNA in maternal plasma from 5 to 41 weeks of gestation
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DOI：
10.1373/clinchem.2004.042713
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发表时间：
2005-02-01
影响因子：
9.3
通讯作者：
Keer, JT
中科院分区：
文献类型：
--
作者：
Birch, L;English, CA;Keer, JT
背景：孕妇血浆中胎儿DNA的检测可以在妊娠5周时实现，但很少有大规模研究报道胎儿和孕妇的DNA在所有三个月内循环。用定量聚合酶链式反应检测总DNA和胎儿DNA浓度，等位基因辨别分析作为特异性检测胎儿DNA的方法。结果：男性胎儿在闭经5周后即可检测到，且胎儿DNA浓度在整个孕期内逐渐升高。经活产确认的胎儿男性性别判定的灵敏度为99%，特异度为100%。总DNA浓度与胎龄无关，但在妊娠早期和晚期略高于中期。对短串联重复序列的分析表明，对胎儿DNA的特异性检测需要显著提高检测下限。结论：基于聚合酶链式反应的检测方法具有很高的灵敏度，结合实时DNA分析提供的定量方法，在无创性产前诊断中具有明显的临床应用潜力。然而，使用最佳匹配数据分析的准确量化、方法标准化和性能控制指标对于稳健的常规非侵入性诊断是必要的。(C)2005年美国临床化学协会。
Background: Detection of fetal DNA in maternal plasma is achievable at 5 weeks of gestation, but few large-scale studies have reported circulating fetal and maternal DNA across all trimesters.Methods: Blood samples were collected from 201 women between 5 and 41 weeks of pregnancy. Quantitative PCR was used to assess total and fetal DNA concentrations, and allelic discrimination analysis was investigated as a route to detecting specifically fetal DNA.Results: Male fetuses were detectable from 5 weeks amenorrhea with increasing fetal DNA concentrations across gestation. The sensitivity of fetal male gender determination in pregnancies with live birth confirmation was 99%, with 100% specificity. Total DNA concentrations did not correlate with gestational age, but appeared slightly higher in the first and third trimesters than in mid-pregnancy. Analysis of short tandem repeats demonstrated that significant improvements in the detection limit are required for specific detection of fetal DNA.Conclusions: The high sensitivity of PCR-based detection, together with quantification provided by real-time DNA analysis, has clear potential for clinical application in noninvasive prenatal diagnosis. However, accurate quantification using best-fit data analysis, standardization of methods, and performance control indicators are necessary for robust routine noninvasive diagnostics. (C) 2005 American Association for Clinical Chemistry.