二代测序在儿童线粒体病诊断中的应用  

Diagnosis of mitochondrial disorders in children with next generation sequencing

作　　者：刘志梅[1] 方方[1] 丁昌红[1] 张炜华[1] 李久伟[1] 杨欣英[1] 王晓慧[1] 伍妘[1] 王红梅[1] 刘丽英[1] 韩彤立[1] 王旭[1] 陈春红[1] 吕俊兰[1] 吴沪生[1] 

机构地区：[1]首都医科大学附属北京儿童医院神经内科,100045

出　　处：《中华儿科杂志》2015年第10期747-753,共7页Chinese Journal of Pediatrics

摘　　要：目的 探讨二代测序在儿童线粒体病诊断中的应用价值.方法 对2012年10月至2014年2月在首都医科大学附属北京儿童医院神经内科就诊患儿根据线粒体病标准进行评分,对70例（男38例、女32例,就诊年龄为3个月至14岁）疑似线粒体病的患儿,应用二代测序进行线粒体基因全测序和（或）与线粒体结构和功能相关的核基因测序,总结和分析基因确诊病例的临床资料.结果 70例疑似线粒体病患儿中,发现线粒体基因和核基因变异21例,其中10例为线粒体基因变异、11例为核基因变异.21例中1例为非线粒体病,由CHAT基因纯合突变（p.I187T）导致的先天性肌无力综合征伴发作性呼吸暂停（CMS-EA）.20例为线粒体病,其中,Leigh综合征10例,线粒体脑肌病伴乳酸酸中毒和卒中样发作综合征4例,Leber遗传性视神经病（LHON）及LHON plus 3例,线粒体DNA耗竭综合征2例,不能分类1例.10例线粒体基因变异均为点突变,分别为A3243G、G3460A、G11778A、T14484C、T14502C、T14487C.10例核基因变异线粒体病患儿中发现5个基因变异,分别为SURF1、PDHA1、NDUFV1、SUCLA2、SUCLG1,共有14种变异类型,其中7种变异类型为未报道的新突变.结论 Leigh综合征是儿童线粒体病中最常见的综合征,应用二代测序不仅能提高Leigh综合征的分子诊断水平,还可早期诊断表现不典型的线粒体综合征以及罕见的线粒体病.Objective To explore the application value of next generation sequencing （NGS） in the diagnosis of mitochondrial disorders.Method According to mitochondrial disease criteria,genomic DNA was extracted using standard procedure from peripheral venous blood of patients with suspected mitochondrial disease collected from neurological department of Beijing Children＇s Hospital Affiliated to Capital Medical University between October 2012 and February 2014.Targeted NGS to capture and sequence the entire mtDNA and exons of the 1 000 nuclear genes related to mitochondrial structure and function.Clinical data were collected from patients diagnosed at a molecular level,then clinical features and the relationship between genotype and phenotype were analyzed.Result Mutation was detected in 21 of 70 patients with suspected mitochondrial disease,in whom 10 harbored mtDNA mutation,while 11 nuclear DNA （nDNA） mutation.In 21 patients,1 was diagnosed congenital myasthenic syndrome with episodic apnea due to CHAT gene p.I187T homozygous mutation,and 20 were diagnosed mitochondrial disease,in which 10 were Leigh syndrome,4 were mitochondrial encephalomyopathy with lactic acidosis and stroke like episodes syndrome,3 were Leber hereditary optic neuropathy （LHON） and LHON plus,2 were mitochondrial DNA depletion syndrome and 1 was unknown.All the mtDNA mutations were point mutations,which contained A3243G,G3460A,G11778A,T14484C,T14502C and T14487C.Ten mitochondrial disease patients harbored homozygous or compound heterozygous mutations in 5 genes previously shown to cause disease：SURF1,PDHA1,NDUFV1,SUCLA2 and SUCLG1,which had 14 mutations,and 7 of the 14 mutations have not been reported.Conclusion NGS has a certain application value in the diagnosis of mitochondrial diseases,especially in Leigh syndrome atypical mitochondrial syndrome and rare mitochondrial disorders.

关 键 词：线粒体疾病 高通量核苷酸测序 基因 线粒体 核基因 

分 类 号：R725.9[医药卫生—儿科] R741[医药卫生—临床医学]