目的:在先天性溶血性贫血(CHA)中,并不总是可以通过评估临床表现和常规实验室检查来确定具体诊断。这项研究的目的是评估下一代测序(NGS)和基于临床外显子组的拷贝数变异(CNV)分析在CHA患者中的实用性。
方法:本研究纳入了来自115个无关家庭的143例CHA病例,进行分子分析。分子分析使用两个不同的临床外显子组对130例患者进行,和9名患者的全外显子组测序。在92例NGS数据中,基于外显子组的CNV调用被整合到传统的单核苷酸变体和小的插入/缺失分析管道中。同一个家庭的四个病人,PK吉普赛变异体采用长程聚合酶链反应进行研究.
结果:在86%的研究组中建立了分子诊断。最常见的突变基因是SPTB(31.7%)和PKLR(28.5%)。对92例病例的CNV分析显示,3例患者在SPTB中有不同大小的大缺失,6例患者在PKLR中有缺失。
结论:在这项研究中,NGS在罕见CHA病例中提供了较高的分子诊断率。CNV的分析有助于诊断成功。
OBJECTIVE: In congenital hemolytic anemias (CHA), it is not always possible to determine the specific diagnosis by evaluating clinical findings and conventional laboratory tests. The aim of this study is to evaluate the utility of next-generation sequencing (NGS) and clinical-exome-based copy number variant (CNV) analysis in patients with CHA.
METHODS: One hundred and forty-three CHA cases from 115 unrelated families referred for molecular analysis were enrolled in the study. Molecular analysis was performed using two different clinical exome panels in 130 patients, and whole-exome sequencing in nine patients. Exome-based CNV calling was incorporated into the traditional single-nucleotide variant and small insertion/deletion analysis pipeline for NGS data in 92 cases. In four patients from the same family, the PK Gypsy variant was investigated using long-range polymerase chain reaction.
RESULTS: Molecular diagnosis was established in 86% of the study group. The most frequently mutated genes were SPTB (31.7%) and PKLR (28.5%). CNV analysis of 92 cases revealed that three patients had different sizes of large deletions in the SPTB and six patients had a deletion in the PKLR.
CONCLUSIONS: In this study, NGS provided a high molecular diagnostic rate in cases with rare CHA. Analysis of the CNVs contributed to the diagnostic success.