Validation for Clinical Use of, and Initial Clinical Experience with, a Novel Approach to Population-Based Carrier Screening using High-Throughput, Next-Generation DNA Sequencing
Validation for Clinical Use of, and Initial Clinical Experience with, a Novel Approach to Population-Based Carrier Screening using High-Throughput, Next-Generation DNA Sequencing
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DOI：
10.1016/j.jmoldx.2013.10.006
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发表时间：
2014-03-01
影响因子：
4.1
通讯作者：
Kennedy, Caleb J.
中科院分区：
文献类型：
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作者：
Hallam, Stephanie;Nelson, Heather;Kennedy, Caleb J.
传统的载体筛选试验设计为仅寻找基因内最常见的突变，这是出于成本考虑。虽然这可以在特定人群中产生特定基因的高检出率（例如高加索人的囊性纤维化），但对于其他种族或混合或未知种族背景的患者来说，它们是次优的。下一代DNA测序提供了一个机会，使用更全面的突变面板提供载体筛选，主要是由检测到的序列变化的临床影响的信息限制。我们描述了一种新一代的基于DNA测序的检测方法，能够以及时和全面的方式可靠地筛查患者样本。在研究环境中的分析准确性已被记录在案。在此，我们描述了为确保我们的检测试剂盒用于临床实践的准确性（分析有效性）和耐用性而进行的其他研究，并提供了我们提供该检测的经验数据。我们使用这种方法筛选11，691例体外受精患者的临床经验已经确定了449个突变等位基因：447个在携带者中，2个在受影响的个体中。我们总共在14个不同的基因中发现了87个不同的突变。发现的突变中约有四分之一不包括在传统的、有限的突变组中，包括16个我们小组独有的已知突变，以及几个基因中的新型截短突变。
Traditional carrier screening assays are designed to look for only the most common mutations within a gene owing to cost considerations. Although this can yield high detection rates in specific populations for specific genes (such as cystic fibrosis in Caucasians), they are suboptimal for other ethnicities or for patients of mixed or unknown ethnic background. Next-generation DNA sequencing provides an opportunity to provide carrier screening using more comprehensive mutation panels that are Limited primarily by information about the clinical impact of detected sequence changes. We describe a next-generation DNA sequencing-based assay capable of reliably screening patient samples in a timely and comprehensive manner. The analytic accuracy in a research setting has been documented. Here, we describe the additional studies performed to ensure the accuracy (analytic validity) and robustness of our assay for use in clinical practice and provide data from our experience offering this testing. Our clinical experience using this approach to screen 11,691 in vitro fertilization patients has identified 449 mutant alleles: 447 in carriers and 2 in an affected individual. In total, we found 87 distinct mutations in 14 different genes. Approximately one quarter of the mutations found are not included in traditional, Limited, mutation panels, including 16 known mutations unique to our panel, and novel truncating mutations in several genes.