【作者】 陈茜；

【导师】 傅立军；

【作者基本信息】 上海交通大学 ， 儿科学， 2017， 硕士

【摘要】 目的:分析上海儿童医学中心诊治的婴儿型Pompe病(infantile-onset Pompe disease,IOPD)患儿的临床资料,探讨中国大陆地区IOPD的临床特点分子遗传学特点及其预后,为IOPD的早期诊断、合理治疗及分子诊断和遗传咨询提供依据。方法:回顾性分析2013年至2016年上海儿童医学中心确诊的25例IOPD患儿的病史、首发症状、体征、生化检查结果及心脏超声等辅助检查结果,并对患儿的生存情况等随访资料进行分析。设计GAA基因PCR扩增引物,采用Taq DNA聚合酶试剂盒对DNA样本GAA基因(外显子及其侧翼序列)扩增,扩增后电泳目标条带特异,ABI3100测序仪Sanger测序。结果:25例患儿通过临床表现、外周血GAA酶活性检测及基因检测确诊为IOPD,其中男14例,女11例。其中23例(92%)有P-R间期缩短表现;25例患儿的超声心动图均提示有心肌肥厚,左心室质量指数为133-575g/m~2。GAA基因检测共发现30种不同的突变位点,其中13种为新发现的突变。在本组IOPD患儿中发现两个热点突变,无义突变c.2662G>T(p.Glu888X)为最常见的突变位点,占该组检测到的突变等位基因总数的14%;错义突变c.1935C>A(p.Asp645Glu)是本研究中第二常见的突变位点,占该组检测到的突变等位基因总数的12%。结论:IOPD是一种快速致死性的疾病,心肌肥厚以及肌力/肌张力减退是IOPD最突出的表现。我们在25例中国大陆地区IOPD患者中发现了13个新的突变位点和2个热点突变,c.2662G>T为中国北方地区IOPD患儿最常见的突变,c.1935C>A为中国南方地区IOPD患儿最常见的突变。更多还原

【Abstract】 Objective: To better understand the clinical course and prognosis of infantile-onset Pompe disease(IOPD)in China’s mainland,we analyze the clinical data and explore the molecular genetic characterizations of IOPD patients in Shanghai Children’s Medical Center.In this way,we provide evidences of early diagnosis,reasonable treatment,molecular diagnosis and genetic counseling of IOPD.Methods: Twenty-five Chinese patients with IOPD diagnosed by Shanghai Children’s Medical Center from 2013 to 2016 were enrolled in this study.Their clinical data of initial presentation,sign,laboratory tests and auxiliary examinations,such as X-ray,electrocardiogram and heart ultrasound,were retropspectively reviewed.Their motor development,child survival and other follow-up data were evaluated.Peripheral blood(2ml)was collected from all the individuals and genomic DNA was extracted using a commercial kit..All coding exons(e2-20)and intron/exon boundaries of the GAA gene were amplified by polymerase chain reaction using primers with 16 pairs of primers.The PCR products were analyzed by forward and reverse sequencing.The resulting DNA was sequenced analyzed using the forward and reverse primers.All the mutations were sequence analyzed with their parents.Each novel mutation was screened in 60 normal individuals using direct sequencing.Results: A total of 25 unrelated patients(14 males and 11 females)were diagnosed with IOPD in our institution based on the clinical presentations,GAA enzyme activity assay and gene testing.23(92%)patients had shortened P-R interval performance.All the 25 children were complicated with cardiac hypertrophy,and the left ventricular mass index(LVMI)was 133-575g/m2.A total of 30 different mutations were identified in this study,and 13 of which were novel.Two hot spots were found in children with IPOD.Nonsense mutation c.2662G> T(p.Glu888X)is the most common mutation identified in this study.This mutant allele accounts for an average of 14% of total mutant alleles detected in this group.The missense mutation c.1935C>A(p.Asp645Glu)was the next most common mutation identified in this study.This mutant allele accounts for an average of 12% of total mutant alleles detected in this group.Conclusion: IOPD is a progressive and often fatal disease.Hypertrophic cardiomyopathy,hypotonia,muscle weakness were reported as frequently presenting signs and symptoms in patients with IOPD.We found 13 new mutations and 2 hotspots in 25 patients with IOPD in China’s mainland.c.2662G> T is the most common mutation in children with IPOD in northern China,whereas c.1935C> A is the most common mutation in children with IPOD in southern China.更多还原