PDF(Pubmed)
Abstract：
Massive parallel sequencing methods, such as exome, genome, and targeted DNA sequencing, have aided molecular diagnosis of genetic diseases in the last 20 years. However, short-read sequencing methods still have several limitations, such inaccurate genome assembly, the inability to detect large structural variants, and variants located in hard-to-sequence regions like highly repetitive areas. The recently emerged PacBio single-molecule real-time (SMRT) and Oxford nanopore technology (ONT) long-read sequencing (LRS) methods have been shown to overcome most of these technical issues, leading to an increase in diagnostic rate. LRS methods are contributing to the detection of repeat expansions in novel disease-causing genes (e.g., ABCD3, NOTCH2NLC and RILPL1 causing an Oculopharyngodistal myopathy or PLIN4 causing a Myopathy with rimmed ubiquitin-positive autophagic vacuolation), of structural variants (e.g., in DMD), and of single nucleotide variants in repetitive regions (TTN and NEB). Moreover, these methods have simplified the characterization of the D4Z4 repeats in DUX4, facilitating the diagnosis of Facioscapulohumeral muscular dystrophy (FSHD). We review recent studies that have used either ONT or PacBio SMRT sequencing methods and discuss different types of variants that have been detected using these approaches in individuals with neuromuscular disorders.
摘要：
大规模平行测序方法，比如exome，基因组,和靶向DNA测序，在过去的20年里帮助了遗传疾病的分子诊断。然而,短读测序方法仍然有一些局限性，如此不准确的基因组组装，无法检测到大型结构变体，和位于难以测序区域的变异体，如高度重复的区域。最近出现的PacBio单分子实时（SMRT）和牛津纳米孔技术（ONT）长读取测序（LRS）方法已被证明可以克服大多数这些技术问题，导致诊断率的提高。LRS方法有助于检测新的致病基因中的重复扩增(例如，ABCD3，NOTCH2NLC和RILPL1引起耳咽远端肌病或PLIN4引起带泛素阳性自噬空泡的肌病），结构变体(例如，在DMD中)，以及重复区（TTN和NEB）中的单核苷酸变体。此外，这些方法简化了DUX4中D4Z4重复序列的表征,有助于面肩肱型肌营养不良(FSHD)的诊断.我们回顾了最近使用ONT或PacBioSMRT测序方法的研究，并讨论了使用这些方法在神经肌肉疾病患者中检测到的不同类型的变异。
