A high-throughput sequencing test for diagnosing inherited bleeding, thrombotic, and platelet disorders
A high-throughput sequencing test for diagnosing inherited bleeding, thrombotic, and platelet disorders
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DOI：
10.1182/blood-2015-12-688267
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发表时间：
2016-06-09
期刊：
影响因子：
20.3
通讯作者：
Turro, Ernest
中科院分区：
文献类型：
--
作者：
Simeoni, Ilenia;Stephens, Jonathan C.;Turro, Ernest
遗传性出血、血栓形成和血小板疾病（BPD）是每百万例新生儿中影响约300例的疾病。除血友病和血管性血友病患者外，通常无法对BPD患者进行分子分析。通常需要许多专门的测试来达到推定的诊断，并且通常以逐步的方式进行以控制成本。这种方法会导致延误，并且通常无法达到结论性的分子诊断，这可能会影响治疗并阻碍受影响亲属的快速识别。为了解决这一未满足的诊断需求，我们设计了一个高通量测序平台，靶向与BPD相关的63个基因。该平台可以调用单核苷酸变异，短插入/缺失和大拷贝数变异（尽管不是倒位），这些变异经过自动过滤以进行诊断优先级排序，导致每个个体平均5.34个候选变异。我们分别对159和137个样本进行了测序，这些样本来自有和没有先前已知的因果变异的病例。在后一组中，61例病例的临床和实验室表型表明一个特定的分子病因，而其余的先验高度不确定的病因。所有以前检测到的变异被概括，当病因被怀疑，但未知或不确定，分子诊断达到56 61例和76例中只有8例，分别。后一类突出了需要进一步研究BPD的新原因。因此，ThromboGenomics平台提供了一种负担得起的基于DNA的测试，以诊断疑似患有已知遗传性BPD的患者。
Inherited bleeding, thrombotic, and platelet disorders (BPDs) are diseases that affect similar to 300 individuals per million births. With the exception of hemophilia and von Willebrand disease patients, a molecular analysis for patients with a BPD is often unavailable. Many specialized tests are usually required to reach a putative diagnosis and they are typically performed in a step-wise manner to control costs. This approach causes delays and a conclusive molecular diagnosis is often never reached, which can compromise treatment and impede rapid identification of affected relatives. To address this unmet diagnostic need, we designed a high-throughput sequencing platform targeting 63 genes relevant for BPDs. The platform can call single nucleotide variants, short insertions/deletions, and large copy number variants (though not inversions) which are subjected to automated filtering for diagnostic prioritization, resulting in an average of 5.34 candidate variants per individual. We sequenced 159 and 137 samples, respectively, from cases with and without previously known causal variants. Among the latter group, 61 cases had clinical and laboratory phenotypes indicative of a particular molecular etiology, whereas the remainder had an a priori highly uncertain etiology. All previously detected variants were recapitulated and, when the etiology was suspected but unknown or uncertain, a molecular diagnosis was reached in 56 of 61 and only 8 of 76 cases, respectively. The latter category highlights the need for further research into novel causes of BPDs. The ThromboGenomics platform thus provides an affordable DNA-based test to diagnose patients suspected of having a known inherited BPD.