MRC Clinical Sciences Centre, Imperial College London, London, UK.
J Cardiovasc Transl Res. 2013 Feb;6(1):94-103. doi: 10.1007/s12265-012-9401-8. Epub 2012 Sep 7.
Next-generation sequencing (NGS) provides an unprecedented opportunity to assess genetic variation underlying human disease. Here, we compared two NGS approaches for diagnostic sequencing in inherited arrhythmia syndromes. We compared PCR-based target enrichment and long-read sequencing (PCR-LR) with in-solution hybridization-based enrichment and short-read sequencing (Hyb-SR). The PCR-LR assay comprehensively assessed five long-QT genes routinely sequenced in diagnostic laboratories and "hot spots" in RYR2. The Hyb-SR assay targeted 49 genes, including those in the PCR-LR assay. The sensitivity for detection of control variants did not differ between approaches. In both assays, the major limitation was upstream target capture, particular in regions of extreme GC content. These initial experiences with NGS cardiovascular diagnostics achieved up to 89 % sensitivity at a fraction of current costs. In the next iteration of these assays we anticipate sensitivity above 97 % for all LQT genes. NGS assays will soon replace conventional sequencing for LQT diagnostics and molecular pathology.
下一代测序（NGS）为评估人类疾病相关的遗传变异提供了前所未有的机会。在这里，我们比较了两种用于遗传性心律失常综合征诊断测序的 NGS 方法。我们比较了基于 PCR 的靶向富集和长读测序（PCR-LR）与基于溶液杂交的富集和短读测序（Hyb-SR）。PCR-LR 检测全面评估了五个在诊断实验室常规测序的长 QT 基因和 RYR2 中的“热点”。Hyb-SR 检测靶向 49 个基因，包括 PCR-LR 检测中的基因。检测对照变异的敏感性在两种方法之间没有差异。在两种检测方法中，主要的限制是上游靶标捕获，特别是在极端 GC 含量的区域。这些 NGS 心血管诊断的初步经验以当前成本的一小部分实现了高达 89%的敏感性。在这些检测方法的下一个迭代中，我们预计所有 LQT 基因的敏感性都将超过 97%。NGS 检测方法将很快取代传统的 LQT 诊断和分子病理学测序。
