Adapting SureSelect enrichment protocol to the Ion Torrent S5 platform in molecular diagnostics of craniosynostosis
Adapting SureSelect enrichment protocol to the Ion Torrent S5 platform in molecular diagnostics of craniosynostosis
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DOI：
10.1038/s41598-020-61048-5
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发表时间：
2020-03-05
影响因子：
4.6
通讯作者：
Jamsheer, Aleksander
中科院分区：
文献类型：
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作者：
Bukowska-Olech, Ewelina;Popiel, Delfina;Jamsheer, Aleksander
获得可靠和高保真的下一代测序（NGS）数据需要选择合适的测序平台和文库制备方法，这两者都有其固有的分析特异性局限性。在这里，我们展示了成功适应基于SureSelect杂交的靶标富集协议的结果，该协议在Ion Torrent S5平台上测序，该平台旨在更好地与基于扩增子的面板一起工作。在我们的研究中，我们应用了定制的NGS面板来筛选16名不相关的颅缝过早融合（即颅缝闭合（CS））患者。CS要么作为孤立的畸形，要么以综合征的形式出现，代表了一组遗传异质性和临床可变的疾病。本文提出的方法使我们能够在19%的病例中获得高质量的NGS数据和确诊的分子诊断，达到与一些已发表的研究报告相似的诊断率。总之，我们证明了一种替代富集策略可以成功地在Ion Torrent S5平台上测序之前应用于文库制备。同时，我们也证明了我们设计的定制NGS面板在CS患者的分子诊断中是一个有用和有效的工具。
Obtaining reliable and high fidelity next-generation sequencing (NGS) data requires to choose a suitable sequencing platform and a library preparation approach, which both have their inherent assay-specific limitations. Here, we present the results of successful adaptation of SureSelect hybridisation-based target enrichment protocol for the sequencing on the Ion Torrent S5 platform, which is designed to work preferably with amplicon-based panels. In our study, we applied a custom NGS panel to screen a cohort of 16 unrelated patients affected by premature fusion of the cranial sutures, i.e. craniosynostosis (CS). CS occurs either as an isolated malformation or in a syndromic form, representing a genetically heterogeneous and clinically variable group of disorders. The approach presented here allowed us to achieve high quality NGS data and confirmed molecular diagnosis in 19% of cases, reaching the diagnostic yield similar to some of the published research reports. In conclusion, we demonstrated that an alternative enrichment strategy for library preparations can be successfully applied prior to sequencing on the Ion Torrent S5 platform. Also, we proved that the custom NGS panel designed by us represents a useful and effective tool in the molecular diagnostics of patients with CS.