Non-invasive prenatal diagnosis of achondroplasia and thanatophoric dysplasia: next-generation sequencing allows for a safer, more accurate, and comprehensive approach.
Non-invasive prenatal diagnosis of achondroplasia and thanatophoric dysplasia: next-generation sequencing allows for a safer, more accurate, and comprehensive approach.
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DOI：
10.1002/pd.4583
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发表时间：
2015-07
影响因子：
3
通讯作者：
Jenkins LA
中科院分区：
文献类型：
--
作者：
Chitty LS;Mason S;Barrett AN;McKay F;Lench N;Daley R;Jenkins LA
准确的产前诊断遗传疾病可能具有挑战性，通常需要侵入性检测。在这里，我们展示了下一代测序（NGS）的潜力，用于分析母体血液中的游离DNA，以改变单基因疾病的产前诊断。在有软骨发育不全和致死性发育不良风险的孕妇中，使用PCR和限制性酶消化（PCR-RED）分析游离DNA与新的NGS测定进行比较。PCR-RED检测72例，正确率88.6%，不确定率7%，假阴性1例。NGS检查47例，准确率96.2%，无无效病例。两种方法都用于27例病例，其中NGS在PCR-RED不确定的两例病例中给出了正确的结果。NGS为新生和父系遗传突变的非侵入性产前诊断提供了一种准确、灵活的方法。它比PCR-RED更敏感，在筛选具有多个潜在致病突变的基因时是理想的。这些发现突出了NGS在其他单基因疾病的非侵入性产前诊断中的价值。© 2015作者。Prenatal Diagnosis由John Wiley & Sons，Ltd.出版。据报道，使用基于PCR的方法进行的非侵入性产前诊断（NIPD）可用于检测或排除母体血浆中的个体父系遗传或从头等位基因。使用下一代测序的NIPD提供了一种准确、更灵敏的方法，可用于在单次测定中检测多个突变，因此在筛选具有多个潜在致病突变的基因时是理想的。因此，下一代测序为非侵入性产前诊断提供了一种灵活的方法，非常适合在忙碌的服务实验室中使用。
Accurate prenatal diagnosis of genetic conditions can be challenging and usually requires invasive testing. Here, we demonstrate the potential of next-generation sequencing (NGS) for the analysis of cell-free DNA in maternal blood to transform prenatal diagnosis of monogenic disorders. Analysis of cell-free DNA using a PCR and restriction enzyme digest (PCR–RED) was compared with a novel NGS assay in pregnancies at risk of achondroplasia and thanatophoric dysplasia. PCR–RED was performed in 72 cases and was correct in 88.6%, inconclusive in 7% with one false negative. NGS was performed in 47 cases and was accurate in 96.2% with no inconclusives. Both approaches were used in 27 cases, with NGS giving the correct result in the two cases inconclusive with PCR–RED. NGS provides an accurate, flexible approach to non-invasive prenatal diagnosis of de novo and paternally inherited mutations. It is more sensitive than PCR–RED and is ideal when screening a gene with multiple potential pathogenic mutations. These findings highlight the value of NGS in the development of non-invasive prenatal diagnosis for other monogenic disorders. © 2015 The Authors. Prenatal Diagnosis published by John Wiley & Sons, Ltd. Non-invasive prenatal diagnosis (NIPD) using PCR-based methods has been reported for the detection or exclusion of individual paternally inherited or de novo alleles in maternal plasma. NIPD using next generation sequencing provides an accurate, more sensitive approach which can be used to detect multiple mutations in a single assay and so is ideal when screening a gene with multiple potential pathogenic mutations. Next generation sequencing thus provides a flexible approach to non-invasive prenatal diagnosis ideal for use in a busy service laboratory.