张林妹　周渊峰　柴毅明　王　佶　吴冰冰　王　艺　周水珍



·论著·

结节性硬化105例临床特征和基因型分析

张林妹周渊峰柴毅明王佶吴冰冰王艺周水珍

结节性硬化；诊断标准；临床特征；TSC基因

AbstractObjectiveTo analyse the clinical phenotype and genotype of tuberous sclerosis complex (TSC) and improve the diagnostic level of TSC.MethodsPatients were retrospectively collected from Aug. 2013 to Sept. 2015 hospitalized in Children′s Hospital of Fudan University, who were diagnosed with 2012 revised diagnostic criteria for TSC. Data of clinical phenotype included brain, skin, heart, kidney and eyes were intercepted. Patients were divided into the age of <1 year, -3 years, -6 years, -13 years and -18 years groups, statistics was made to get detection rate and the correlation was analysed between genetic mutation and clinical phenotype.ResultsA total of 105 cases diagnosed as TSC were recruited into the study including 54 males and 51 females. Age of visit was ranged from 2 months to 13 years. ① Reason for the first visit: 83 cases (79.0%) for seizure, 8 cases (7.6%) for abnormal of skin and 5 cases (4.8%) for cardiac tumor. ②39 cases (37.1%) were diagnosed only with clinical phenotype; 47(44.8%) with clinical phenotype and genetic testing; 2 cases (1.9%) with clinical suspicious but gene positive. ③Clinical characteristics detection rate: Subependymal nodules were in 91/99 cases(91.9%), cortical dysplasias were in 81/99 cases(81.8%), which was similar with other different age groups. Hypomelanotic macules were in 97/105 cases(92.4%), angiofibromas were in 55 cases(52.4%), shagreen patch in 46 cases(43.8%), the detection rate of angiofibromas and shagreen patch rised with age. Cardiac rhabdomyomas were in 25/75 cases(33.3%)，the detection rate descended with age; renal lesions were in 14/71 cases(19.7%); ocular lesions were in 5/37 cases(13.5%). ④TSC1 mutation was detected in 15/66 cases (22.7%)，TSC2 mutation was detected in 34/66 cases (51.5%)；spasm was more common in patients withTSC2 mutation (29.4%vs13.3%).ConclusionTSC is an extremely variable disease that can affect multiple important organs. The detection rates of brain lesion were similar among different age and cardiac rhabdomyomas decreased with age. Gene test conduced to the diagnosis of clinical suspect cases.

结节性硬化 (TSC) 是一种罕见的常染色体显性遗传疾病[1]。TSC皮肤病变影响美观，且可累及脑、心、肾、肺、肝和眼等重要器官。Gomez于1979年首先提出了TSC诊断标准，1998年国际TSC临床共识会议对该标准进行了修订[2,3]，近年来随着影像学和基因检测技术的进展， 2012年修订了新的TSC诊断标准[1]，其中临床特征仍是TSC诊断的主要方法，补充基因检测结果作为独立的诊断标准，诊断的级别从3个(可疑、可能、确诊)降至2个(可能、确诊)。目前国内尚缺乏应用此标准诊断TSC的大宗病例报道，也缺乏TSC基因型与临床特征的相关性分析。本文回顾性分析复旦大学附属儿科医院(我院)以2012年修订标准诊断的TSC病例的临床特征和基因型，以期提高临床医生对本病的认识。

1　方法

1.2病例纳入标准2013年8月至2015年9月在我院神经科TSC专病门诊就诊的TSC病例，需要说明的是我院2013年8月后执行2012年修订的TSC诊断标准。

1.3TSC基因检测方法采集外周抗凝血2 mL，使用QIAamp DNA Mini Kit抽提基因组DNA，NanoDrop紫外分光光度仪检测DNA浓度和A260/A280比值。参考国内外文献并使用Primer 3在线软件设计TSC1和TSC2基因PCR引物，TaKaRa LA Taq酶进行扩增，2%琼脂糖电泳以判断产物的扩增特异性。参照Ion文库构建相关试剂盒使用说明书制备测序文库，步骤简述如下：将单个样本的PCR产物标准化后全部混合，经酶切打断，接头连接，片段选择，产物定量，乳液PCR后制备模板，磁珠富集纯化。……