Clinical utility of next-generation sequencing in the diagnosis of hereditary haemolytic anaemias
Clinical utility of next-generation sequencing in the diagnosis of hereditary haemolytic anaemias
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DOI：
10.1111/bjh.14131
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发表时间：
2016-09-01
影响因子：
6.5
通讯作者：
Christensen, Robert D.
中科院分区：
文献类型：
--
作者：
Agarwal, Archana M.;Nussenzveig, Roberto H.;Christensen, Robert D.
遗传性溶血性贫血是一种遗传和表型异质性疾病，其特征是红细胞破坏增加，后果从无害到严重危及生命的贫血。诊断实验室致力于通过使用主要基于形态学和生物化学技术的测试来帮助临床医生达到准确的患者诊断。然而，这些常规研究可能是不确定的，特别是在新生儿和最近进行输血时。大量和大小的潜在涉及的基因也强加了一个实际的挑战，分子诊断使用常规测序方法。为了克服这些诊断缺陷，我们利用下一代测序技术提供高通量、高灵敏度的检测。我们开发了一个面板询问28个基因编码的细胞骨架蛋白和酶的编码区，剪接位点连接，深内含子和调控区的测序覆盖。然后，我们评估了19个样本，包括不明原因的极端高胆红素血症的婴儿和输血依赖性溶血性贫血的患者。在可能的情况下，致病性突变的遗传模式通过直系亲属的测序来确定。我们的结论是，这种下一代测序小组可能是遗传性溶血性贫血分子诊断的一种具有成本效益的方法，特别是当家族史信息不足或常规实验室检测未能识别致病性溶血过程时。
Hereditary haemolytic anaemias are genetically and phenotypically heterogeneous disorders characterized by increased red cell destruction, with consequences ranging from innocuous to severe life-threatening anaemia. Diagnostic laboratories endeavour to assist clinicians reach the exact patient diagnosis by using tests principally based on morphological and biochemical techniques. However, these routine studies may be inconclusive, particularly in newborn infants and when transfusions have recently been administered. Large numbers and size of the potentially involved genes also impose a practical challenge for molecular diagnosis using routine sequencing approaches. To overcome these diagnostic shortcomings, we have utilized next-generation sequencing to provide a high-throughput, highly sensitive assay. We developed a panel interrogating 28 genes encoding cytoskeletal proteins and enzymes with sequencing coverage of the coding regions, splice site junctions, deep intronic and regulatory regions. We then evaluated 19 samples, including infants with unexplained extreme hyperbilirubinaemia and patients with transfusion-dependent haemolytic anaemia. Where possible, inheritance patterns of pathogenic mutations were determined by sequencing of immediate relatives. We conclude that this next-generation sequencing panel could be a cost-effective approach to molecular diagnosis of hereditary haemolytic anaemia, especially when the family history is uninformative or when routine laboratory testing fails to identify the causative haemolytic process.