Diagnosis of Primary Ciliary Dyskinesia by a Targeted Next-Generation Sequencing Panel Molecular and Clinical Findings in Italian Patients
Diagnosis of Primary Ciliary Dyskinesia by a Targeted Next-Generation Sequencing Panel Molecular and Clinical Findings in Italian Patients
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DOI：
10.1016/j.jmoldx.2016.07.002
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发表时间：
2016-11-01
影响因子：
4.1
通讯作者：
Vazza, Giovanni
中科院分区：
文献类型：
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作者：
Boaretto, Francesca;Snijders, Deborah;Vazza, Giovanni
原发性纤毛运动障碍(PCD)是一种罕见的遗传性疾病，会改变粘液纤毛的清除，从而导致上呼吸道和下呼吸道的慢性疾病。PCD的诊断具有挑战性，由于在34个不同的基因中发现了常染色体隐性致病突变，因此该疾病的高度异质性阻碍了基因检测。在这项研究中，我们对51名有PCD临床症状的意大利患者进行了临床和分子特征分析。一种定制的下一代测序面板被开发出来，它能够负担得起并同时筛选24个PCD基因，用于基因分析。经过变异筛选和优先排序，43%的患者获得了PCD的分子诊断。总的来说，在11个PCD基因中发现了5个纯合子突变和27个复合杂合子突变，其中21个以前从未报道过。DNAH5和DNAH11基因是意大利PCD最常见的原因，但也发现了一些群体特异性。此外，未解决病例的数量和在6名患者中仅发现一种突变表明了进一步的遗传异质性，并需要新的策略来检测非传统致病DNA变异。最后，尽管有突变数据库和二氧化硅预测工具帮助解释下一代测序筛查中的变异，但需要进行全面的分离分析，以确定反式遗传和支持突变的致病作用。
Primary ciliary dyskinesia (PCD) is a rare genetic disorder that alters mucociliary clearance, with consequent chronic disease of upper and lower airways. Diagnosis of PCD is challenging, and genetic testing is hampered by the high heterogeneity of the disease, because autosomal recessive causative mutations were found in 34 different genes. In this study, we clinically and molecularly characterized a cohort of 51 Italian patients with clinical signs of PCD. A custom next-generation sequencing panel that enables the affordable and simultaneous screening of 24 PCD genes was developed for genetic analysis. After variant filtering and prioritization, the molecular diagnosis of PCD was achieved in 43% of the patients. Overall, 5 homozygous and 27 compound heterozygous mutations, 21 of which were never reported before, were identified in 11 PCD genes. The DNAH 5 and DNAH11 genes were the most common cause of PCD in Italy, but some population specificities were identified. In addition, the number of unsolved cases and the identification of only a single mutation in six patients suggest further genetic heterogeneity and invoke the need of novel strategies to detect unconventional pathogenic DNA variants. Finally, despite the availability of mutation databases and in silica prediction tools helping the interpretation of variants in next-generation sequencing screenings, a comprehensive segregation analysis is required to establish the in trans inheritance and support the pathogenic role of mutations.