用于无创产前诊断的SRY基因高灵敏高特异检测方法研究

发布时间：2018-04-29 11:41

  本文选题：实时荧光PCR + 核酸侵入反应　； 参考：《分析化学》2017年10期

【摘要】：通过检测母体外周血中胎儿游离DNA(cffDNA)的SRY基因,确定胎儿性别,可评估胎儿性连锁遗传病的发病风险,降低病儿出生率。本研究建立了高灵敏、高特异、闭管检测不易污染的实时荧光PCR偶联核酸侵入反应方法用于SRY基因的检测。通过优化反应体系中的检测探针浓度、FEN1酶用量、Taq酶用量及预扩增退火温度,确定了最佳的反应条件,即检测探针浓度为250 nmol/L、FEN1酶用量为7.5 U、Taq酶用量为0.5 U、预扩增退火温度为67℃。在最佳反应条件下,实现对含量低至4‰(4 copies/μL)的模拟样本的检测,并成功检测两例孕期分别为9周和10周的临床实际样本。结果表明,所建立的方法可用于母体外周血cffDNA的SRY基因检测,为临床开展基于SRY基因的无创产前诊断提供了新方法。
[Abstract]:By detecting the SRY gene of fetal free DNA in maternal peripheral blood and determining the fetal sex, we can evaluate the risk of fetal linked genetic disease and reduce the birth rate. In this study, a real-time fluorescent PCR coupled nucleic acid invasion method with high sensitivity, high specificity and low contamination was established for the detection of SRY gene. The optimum reaction conditions were determined by optimizing the probe concentration and the amount of FEN1 enzyme and Taq enzyme and the pre-amplification annealing temperature in the reaction system. The optimum reaction conditions were as follows: the detection probe concentration was 250 nmol / L FERN1 enzyme dosage 7.5 UTaq enzyme dosage was 0.5 UTaq enzyme dosage, the pre-amplification annealing temperature was 67 鈩，

本文编号：1819768