线粒体基因大片段缺失在Kearns-Sayre综合征不同组织中的研究  

Large-single scale mitochondrial DNA deletions in different tissues in Kearns-Sayre syndrome

作　　者：石雨晴 方方[1] 刘志梅[1] 张炜华[1] 李久伟[1] 陈国洪 吕俊兰[1] 丁昌红[1] 任晓暾[1] Shi Yuqing;Fang Fang;Liu Zhimei;Zhang Weihua;Li Jiuwei;Chen Guohong;Lyu Junlan;Ding Changhong;Ren Xiaotun(Department of Neurology,Beijing Children′s Hospital,Capital Medical University,Beijing 100045,China;Department of Neurology,Children′s Hospital Affiliated to Zhengzhou University,Henan Children′s Hospital,Zhengzhou Children′s Hospital Eastern Region,Zhengzhou 450018,China)

机构地区：[1]首都医科大学附属北京儿童医院神经内科,100045 [2]郑州大学附属儿童医院,河南省儿童医院,郑州儿童医院东区神经内科,450018

出　　处：《中华实用儿科临床杂志》2019年第20期1550-1554,共5页Chinese Journal of Applied Clinical Pediatrics

摘　　要：目的利用长片段PCR扩增二代测序技术在儿童Kearns-Sayre综合征(KSS)的外周血、尿液和肌肉组织等不同样本中进行线粒体基因大片段缺失分析,寻找一种无创性的诊断手段。方法收集2016年10月至2017年10月首都医科大学附属北京儿童医院神经内科住院的KSS患儿的外周血、尿液和肌肉标本,利用二代测序技术对线粒体DNA和全外显子组进行检测。收集患儿临床资料、肌肉组织和酶学检测结果。结果符合KSS临床诊断4例,平均发病年龄8.2岁,首发症状分别为身材矮小、眼睑下垂、头痛、呕吐和视力下降。4例均出现眼球活动受限、运动不耐受、生长迟缓(身高、体质量均在健康同龄儿第3百分位以下)、发育迟滞、食欲不振、肌无力、肌张力低下、脑脊液蛋白>1000 mg/L,头颅磁共振成像(MRI)均提示深部灰质核团和周围白质多发对称异常信号及脑萎缩样表现。其中3例有心脏传导系统异常,1例有色素性视网膜变性,2例有母系家族史。4例在外周血、尿液和肌肉标本中均检测到线粒体DNA单一大片段缺失,分别为m.6460-15590范围内9131 bp缺失,m.8482-13446范围内4964 bp缺失,m.6831-14981范围内8151 bp缺失,m.7983-15495范围内7513 bp缺失。其中3例进行了家系分析,1例母亲外周血和尿液中检测到相同异质性缺失变异,余为阴性。结论KSS罕见,该病在外周血、尿液和肌肉组织中均存在线粒体基因大片段缺失,利用长片段PCR二代测序技术可在外周血和尿液中检测到大片段缺失,为KSS患者提供了一种无创的诊断手段。Objective To investigate the clinical significance of different samples(the peripheral blood,urine and skeletal muscle)that could be detected the large-scale single deletions directly by using next-generation sequencing in the diagnosis of Kearns-Sayre syndrome(KSS)by concluding the clinical and genetic features of KSS,in order to explore a non-invasive method for diagnosis.Methods The clinical data,skeletal muscle′s pathology and enzymology and genetic results of individuals with KSS,who were hospitalized from October 2016 to October 2017 in Department of Neurology,Beijing Children′s Hospital,Capital Medical University,were collected.The gene tests were performed by using next generation sequencing technology and long-PCR technology of mitochondrial DNA(mtDNA)and the whole exon in the peripheral blood,urine and skeletal muscle.Results Four patients were all consistent with the diagnosis criteria of KSS,among whom the age of onset was 8.2 years old on average,and the initial symptoms were statue,ptosis,headache and vomiting,and visual impairment.The common symptoms of the 4 cases were ophthalmoplegia,exercise intolerance,development delay,loss of appetite,hypotonia,muscle weakness,with cerebrospinal fluid protein concentration over 1000 mg/L,the cerebral magnetic resonance imaging showed that abnormal signals in the brainstem,in addition,white matter,thalamus,basal ganglia,cerebrum and cerebellum atrophy could be found.Moreover,3 cases had cardiac conduction block.Two cases had maternal family history.Molecular analysis of the 4 cases revealed the large-scale single deletions of mtDNA from the peripheral blood,the urine,the skeletal muscle through the next-generation sequencing,which were m.6460-15590(9131 bp del),m.8482-13446(4964 bp del),m.6831-14981(8151 bp del),m.7983-15495(7513 bp del),respectively.Among 3 cases who did pedigree analysis,only the mother of case 4 was detected with the same variation of the proband.Conclusions KSS is a rare mitochondrial disease,which could be detected with the single lar

关 键 词：KEARNS-SAYRE综合征 线粒体DNA大片段缺失 二代测序 外周血 尿液 

分 类 号：R73[医药卫生—肿瘤]