姚凤霞;睢瑞芳;张为民;戴毅;郭玉璞;
YAO Feng-xia 1 ,SUI Rui-fang 2 ,ZHANG Wei-min 1 ,DAI Yi 3 ,GUO Yu-pu 3 1 Clinical Research Laboratory, 2 Department of Ophthalmology, 3 Department of Neurology,Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100730,China

• 1:中国医学科学院 北京协和医学院 北京协和医院中心实验室
• 2:中国医学科学院 北京协和医学院 北京协和医院眼科
• 3:中国医学科学院 北京协和医学院 北京协和医院神经科

摘要(Abstract)：

目的探讨多重连接探针扩增技术(multiplex ligation-dependent probe amplification,MLPA)用于常见线粒体疾病基因检测的可行性。方法收集2010年5月至2012年8月北京协和医院281例可疑线粒体异常病例,包括神经科存在神经系统损伤的患者233例及眼科疑诊Leber遗传性视神经病变的患者48例,运用MLPA法对常见线粒体疾病突变位点进行检测,并使用线粒体基因序列测定方法进行验证。结果 281例标本中共38例检测到线粒体基因突变,总检出率为13.5%。眼科48例标本中,3460G>A、11778G>A和14484T>C3种突变共检测到19例,检出率为39.6%;神经科233例标本中,3243A>G、8344A>G、8993T>G和大片段缺失共检测到19例,检出率为8.2%。除1例大片段缺失暂无PCR测序验证外,其余MLPA结果均经序列测定验证为相符。结论 MLPA法是一种可行的快速、准确、简便的基因诊断方法,可作为筛查常见线粒体疾病的检测工具,为临床诊断和治疗提供依据。
Objective To investigate the feasibility of multiplex ligation-dependent probe amplification (MLPA) for the molecular diagnosis of mitochondriopathy. Methods Totally 281 patients with suspected mitochondriopathy in Peking Union Medical College Hospital from May 2010 to August 2012 were enrolled in this study. Among them 233 with nervous system damage were from the department of neurology and 48 with suspected Leber hereditary optic neuropathy (LHON) were from the department of ophthalmology. The common mutation sites for mitochondrial disease were detected with MLPA,and the results were verified by mitochondrial DNA (mtDNA) sequencing. Results We found 38 cases carried mtDNA mutations from all 281 cases,and the total detection rate was 13. 5% . Among 48 cases suspected of LHON,19 cases (39. 6%) were found containing 3460G > A,11778G > A or 14484T > C. Among 233 cases suspected of mitochondria related neuropathies,19 cases (8. 2%) were found containing 3243A > G,8344A > G,8993T > G or a large deletion. Except that the large deletion could not be sequenced,the other mutations detected by MLPA were all verified as consistent with gene sequencing results. Conclusions MLPA is a rapid,accurate,and simple method for detecting mtDNA mutation for common mitochondriopathy. It is feasible to be used in clinical diagnostic laboratory.

关键词(KeyWords)： 线粒体疾病;多重连接探针扩增技术;基因突变;Leber遗传性视神经病变;线粒体脑肌病伴乳酸血症和卒中样发作
mitochondriopathy; multiplex ligation-dependent probe amplification; gene mutation; Leber hereditary optic neuropathy; mitochondrial encephalopathy with lactic acidosis and stroke-like episodes

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81000252)

作者(Author): 姚凤霞;睢瑞芳;张为民;戴毅;郭玉璞;
YAO Feng-xia 1 ,SUI Rui-fang 2 ,ZHANG Wei-min 1 ,DAI Yi 3 ,GUO Yu-pu 3 1 Clinical Research Laboratory, 2 Department of Ophthalmology, 3 Department of Neurology,Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100730,China

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