Centre for Medical Research, University of Western Australia, Harry Perkins Institute of Medical Research, QEII Medical Centre, Nedlands, Western Australia, Australia.
School of Biomedical Sciences, University of Western Australia, Perth, Western Australia, Australia.
Ann Clin Transl Neurol. 2020 Mar;7(3):353-362. doi: 10.1002/acn3.51002. Epub 2020 Mar 9.
To develop, test, and iterate a comprehensive neuromuscular targeted gene panel in a national referral center.
We designed two iterations of a comprehensive targeted gene panel for neuromuscular disorders. Version 1 included 336 genes, which was increased to 464 genes in Version 2. Both panels used TargetSeq probe-based hybridization for target enrichment followed by Ion Torrent sequencing. Targeted high-coverage sequencing and analysis was performed on 2249 neurology patients from Australia and New Zealand (1054 Version 1, 1195 Version 2) from 2012 to 2015. No selection criteria were used other than referral from a suitable medical specialist (e.g., neurologist or clinical geneticist). Patients were classified into 15 clinical categories based on the clinical diagnosis from the referring clinician.
Six hundred and sixty-five patients received a genetic diagnosis (30%). Diagnosed patients were significantly younger that undiagnosed patients (26.4 and 32.5 years, respectively; P = 4.6326E-9). The diagnostic success varied markedly between disease categories. Pathogenic variants in 10 genes explained 38% of the disease burden. Unexpected phenotypic expansions were discovered in multiple cases. Triage of unsolved cases for research exome testing led to the discovery of six new disease genes.
A comprehensive targeted diagnostic panel was an effective method for neuromuscular disease diagnosis within the context of an Australasian referral center. Use of smaller disease-specific panels would have precluded diagnosis in many patients and increased cost. Analysis through a centralized laboratory facilitated detection of recurrent, but under-recognized pathogenic variants.
在国家转诊中心开发、测试和迭代全面的神经肌肉靶向基因组合。
我们设计了两个迭代的神经肌肉疾病综合靶向基因组合。版本 1 包括 336 个基因，在版本 2 中增加到 464 个基因。两个组合都使用 TargetSeq 探针杂交进行靶向富集，然后进行 Ion Torrent 测序。对 2012 年至 2015 年间来自澳大利亚和新西兰的 2249 名神经科患者（1054 名版本 1，1195 名版本 2）进行了靶向高覆盖率测序和分析。除了由合适的医学专家（如神经科医生或临床遗传学家）转诊外，没有使用其他选择标准。根据转诊医生的临床诊断，患者被分为 15 个临床类别。
665 名患者接受了基因诊断（30%）。诊断患者明显比未诊断患者年轻（分别为 26.4 岁和 32.5 岁；P = 4.6326E-9）。疾病类别之间的诊断成功率差异显著。10 个基因中的致病性变异解释了 38%的疾病负担。在多个病例中发现了意想不到的表型扩展。对未解决病例进行研究外显子测试的分类导致发现了六个新的疾病基因。
在澳大利亚转诊中心的背景下，全面的靶向诊断组合是神经肌肉疾病诊断的有效方法。使用较小的疾病特异性组合将排除许多患者的诊断，并增加成本。通过中央实验室进行分析有助于发现反复出现但认识不足的致病性变异。
