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目的探讨新一代测序技术(next-generation sequencing,NGS)无创产前筛查FGFR3相关疾病的可行性与准确率。方法 4例软骨发育不全、2例致死性侏儒Ⅰ型胎儿基因组DNA(g DNA)片段化后,以相应母体产后血浆游离DNA为背景,按照10%、6%、3%、1%、0.5%的浓度进行稀释,形成30例阳性模拟样本,建立孕妇血浆游离DNA模型,同时采集13例阴性对照孕妇血浆游离DNA,运用NGS技术分别检测FGFR3致病突变位点,计算检测灵敏度、特异度、阳性预测值、阴性预测值。结果 30例模拟样本中,胎儿g DNA浓度为10%、6%、3%、1%、0.5%时,突变检出例数分别为6/6、6/6、6/6、3/6、1/6,13例阴性突变检出例数为0/13;胎儿g DNA浓度≥3%时,NGS的灵敏度为100%(95%CI:81.5%~100%),特异度为100%(95%CI:75.3%~100%),阳性预测值为100%(95%CI:81.5%~100%),阴性预测值为100%(95%CI:75.3%~100%)。结论胎儿g DNA浓度≥3%时,NGS可以在孕妇血浆游离DNA模型中准确检出胎儿FGFR3基因突变,提示基于NGS技术的无创产前检测在新发突变或父源遗传疾病的产前诊断领域具有一定的应用前景。
Objective To investigate the feasibility and accuracy of non-invasive prenatal screening of FGFR3-associated diseases by next-generation sequencing (NGS). Methods Four cases of achondroplasia were divided into two groups according to 10%, 6%, 3%, 1%, 0.5% Thirty plasma samples were collected and the free DNA of pregnant women was collected. Thirteen plasma samples of negative controls were collected. NGS was used to detect the pathogenicity of FGFR3. The sensitivity, specificity, and positive Predicted value, negative predictive value. Results The results showed that the number of mutations was 6/6, 6/6, 6/6 and 3/6 respectively in 30 samples of simulated fetal gDNA at the concentrations of 10%, 6%, 3%, 1% and 0.5% , 1/6, and 13 cases with negative mutation were detected in 0/13. The sensitivity of NGS was 100% (95% CI: 81.5% -100%) and the specificity was 100% (95% CI: 75.3% -100%). The positive predictive value was 100% (95% CI: 81.5% -100%) and the negative predictive value was 100% (95% CI: 75.3% -100%). Conclusion NGS can accurately detect fetal FGFR3 gene mutation in plasma DNA of pregnant women when fetal g DNA concentration is more than 3%, suggesting that noninvasive prenatal detection based on NGS technique may play an important role in prenatal diagnosis of new mutations or paternal genetic diseases Has a certain application prospects.