摘要：

研究背景 腓骨肌萎缩症存在高度临床和遗传异质性，传统基因检测需对众多候选基因逐一筛查，存在效率低、耗时、费力等局限性。本文旨在探讨目标区域捕获测序技术诊断常染色体隐性遗传性腓骨肌萎缩症的可行性。 方法 采集5 例临床拟诊常染色体隐性遗传性腓骨肌萎缩症患者的临床资料和外周血样本，采用目标区域捕获测序技术筛查腓骨肌萎缩症相关基因突变，Sanger测序对候选变异位点在患者及其父母外周血样本中进行验证。 结果 目标区域捕获测序显示，2 例检出GDAP1 基因复合杂合突变，余3 例未检出致病性突变。经Sanger 测序证实2 例患儿存在GDAP1 基因突变，例1 为GDAP1 基因复合杂合突变c.767A > G（p.His256Arg）和c.866T > A p.Phe289Tyr），其父携带c.866T > A（p.Phe289Tyr）突变，其母携带c.767A > G（p.His256Arg）突变；例2 为GDAP1 基因复合杂合突变c.571C > T（p.Arg191X）和c.589delC（p.Asp198IlefsX8），其父携带c.589delC（p.Asp198IlefsX8）突变，其母携带c.571C >T（p.Arg191X）突变，最终明确诊断为常染色体隐性遗传性腓骨肌萎缩症。 结论 目标区域捕获测序技术是一项高效基因检测方法，适用于常染色体隐性遗传性腓骨肌萎缩症的基因诊断。

关键词: 夏科-马里-图斯病, 基因, 隐性, 突变, 系谱

Abstract:

Background  Owing to the clinical variability and molecular heterogeneity, the traditional Sanger sequencing takes time and effort and is inefficient. In this paper, target region capture sequencing in the diagnosing of autosomal recessive Charcot-Marie-Tooth disease (AR-CMT) is explored.  Methods  Clinical data and peripheral blood sample of 5 clinically suspected AR - CMT patients were collected. Charcot-Marie-Tooth disease (CMT) related gene mutation were detected by target region capture sequencing. The candidate variants were confirmed in the peripheral blood sample of the patients and their parents by Sanger sequencing. Results  Target region capture sequencing revealed that compound heterozygous mutations of GDAP1 gene was found in 2 patients, and was not seen in the other 3 patients. Sanger sequencing revealed that among the 2 patients with mutation of GDAP1 gene, compound heterozygous mutation c.767A > G (p. His256Arg) and c.866T > A (p. Phe289Tyr) were seen in Case 1, whose father carried c.866T > A (p. Phe289Tyr) heterozygous mutation and mother carried c.767A > G (p. His256Arg) heterozygous mutation, while compound heterozygous mutation c.571C > T (p. Arg191X) and c.589delC (p. Asp198IlefsX8) were seen in Case 2, whose father carried c.589delC (p. Asp198IlefsX8) heterozygous mutation and mother carried c.571C > T (p. Arg191X) heterozygous mutation. Therefore, Case 1 and Case 2 were all diagnosed as AR-CMT.  Conclusions  Target region capture sequencing is a rapid and reliable genetic testing method with high efficiency. It is applicable for the diagnosis of AR-CMT.

Key words: Charcot-Marie-Tooth disease, Genes, recessive, Mutation, Pedigree