Detection of new pathogenic mutations in patients with congenital haemolytic anaemia using next-generation sequencing
Detection of new pathogenic mutations in patients with congenital haemolytic anaemia using next-generation sequencing
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DOI：
10.1111/ijlh.12551
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发表时间：
2016-12-01
影响因子：
3
通讯作者：
Ribeiro, M. L.
中科院分区：
文献类型：
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作者：
Del Orbe Barreto, R.;Arrizabalaga, B.;Ribeiro, M. L.
简介：先天性溶血性贫血（CHA）是指一组遗传异质性疾病，主要由编码珠蛋白链、细胞骨架蛋白和红细胞酶的基因改变引起，其准确诊断可能具有传统技术的挑战性。方法：为了建立一种全面的检测突变的方法，以提高病因学诊断，我们设计了一个定制的面板，使用Ion Torrent (TM) （Thermo Fisher Scientific， S.L. Waltham， MA， USA）个人基因组机（PGM）测序仪，对已知参与CHA发病机制的40个基因的编码区进行测序。对照组包括16个已知突变的样本，试验组包括10个未知突变的患者，分别用于检测验证和应用。结果：在测试组中，我们发现了所有病例的致病突变：4例患者在膜缺陷相关基因（SPTB、ANK1、SLC4A1和EPB41）中出现了新的突变，4例患者在酶缺乏症相关基因（GPI、TPI1和GSS）中出现了纯合或复合杂合突变，1例患者在HBB基因中发生了突变，1例患者在ADAMTS13基因中出现了纯合突变。结论：离子PGM测序与我们的定制面板是检测导致溶血性贫血的突变的高效方法，包括新的变异。这是一种高通量的检测方法，可以在临床实验室中应用。
Introduction: Congenital haemolytic anaemia (CHA) refers to a group of genetically heterogeneous disorders, mainly caused by changes in genes encoding globin chains, cytoskeletal proteins and red cell enzymes, in which accurate diagnosis can be challenging with conventional techniques.Methods: To set-up a comprehensive assay for detecting mutations that could improve aetiological diagnosis, we designed a custom panel for sequencing coding regions from 40 genes known to be involved in the pathogenesis of CHA, using the Ion Torrent (TM) (Thermo Fisher Scientific, S.L. Waltham, MA, USA) Personal Genome Machine (PGM) Sequencer. A control group of 16 samples with previously known mutations and a test group of 10 patients with unknown mutations were included for assay validation and application, respectively.Results: In the test group, we identified pathogenic mutations in all cases: four patients had novel mutations in genes related to membrane defects (SPTB, ANK1, SLC4A1 and EPB41), four were homozygous or compound heterozygous for mutations in genes related to enzyme deficiencies (GPI, TPI1 and GSS), one had a mutation in the HBB gene and another presented a homozygous mutation in the ADAMTS13 gene.Conclusions: Ion PGM sequencing with our custom panel is a highly efficient way to detect mutations causing haemolytic anaemia, including new variations. It is a high-throughput detection method that is ready for application in clinical laboratories.