本文選題：遺傳性痙攣性截癱 + 全外顯子測序��； 參考：《青島大學》2017年碩士論文

【摘要】：目的遺傳性痙攣性截癱(Hereditary Spastic Paraplegia,HSP)是一種具有高度臨床和遺傳異質(zhì)性的神經(jīng)退行性疾病,典型臨床癥狀是雙下肢進行性肌肉緊張、肌無力和痙攣狀態(tài)。本研究旨在應用全外顯子測序?qū)σ粋�中國漢族HSP家系的致病基因進行研究,為家系患者做出準確的基因診斷,為產(chǎn)前診斷和HSP致病機理的研究奠定基礎(chǔ)。方法收集一個包含5代32人的HSP家系,繪制標準家系圖,選取該家系Ⅲ代7人、Ⅳ代12人作為研究對象,采集外周抗凝血樣并提取基因組DNA(g DNA)。對先證者(Ⅳ11)、先證者患病姐姐(Ⅳ8)和先證者未患病哥哥(Ⅳ9)進行一系列臨床檢查,包括病史采集、體格檢查、顱腦磁共振檢查。選�、�11、Ⅳ8和Ⅳ9的g DNA樣本進行全外顯子測序,運用SIFT、Poly Phen2、DNAMAN等生物信息學分析軟件獲取可疑致病基因突變位點。應用PCR擴增及Sanger測序的方法將可疑致病突變位點在該家系19名研究對象和50例與該家系無血緣關(guān)系的健康對照者中進行驗證,分析該位點在家系內(nèi)是否呈共分離,從而進一步驗證該位點與該家系的關(guān)聯(lián)性。結(jié)果臨床檢查結(jié)果:Ⅳ11和Ⅳ8均表現(xiàn)典型雙下肢進行性痙攣性截癱,無脊髓外其他系統(tǒng)癥狀,顱腦、頸椎、胸椎核磁共振檢查均未見明顯異常。基因檢測結(jié)果:通過全外顯子測序和Sanger測序驗證,發(fā)現(xiàn)該家系中6例患者均攜帶SPAST(SPG4)上的同一突變位點(c.1606CT,p.Gln536X),家系中13例正常者和50例與該家系無血緣關(guān)系的健康對照者中均未發(fā)現(xiàn)該突變,表明這一突變位點在該家系中呈共分離。此外,該突變位點在國內(nèi)外尚未報道,是一個新發(fā)現(xiàn)突變位點。結(jié)論結(jié)合臨床癥狀和基因檢測結(jié)果確診先證者及家系其他患者所患疾病為單純型遺傳性痙攣性截癱,新發(fā)現(xiàn)的SPAST(SPG4)上c.1606CT,p.Gln536X雜合突變是該HSP家系的致病基因突變,這不僅拓寬了SPAST(SPG4)基因突變譜,也為該家系產(chǎn)前診斷提供了依據(jù),為研究HSP的發(fā)病機制奠定了理論基礎(chǔ)。
[Abstract]:Objective hereditary spastic paraplegia (Spastic) is a neurodegenerative disease with high clinical and genetic heterogeneity. The typical clinical symptoms are progressive muscle tension, myasthenia and spasm of the lower extremities. The purpose of this study was to study the pathogenicity genes of a Chinese Han HSP family by using total exon sequencing, and to provide a basis for the prenatal diagnosis and the study of the pathogenesis of HSP in pedigrees. Methods A HSP pedigree consisting of 32 persons of 5 generations was collected, and the standard family map was drawn. Seven members of the third generation and 12 people of the fourth generation were selected as the research objects. Peripheral anticoagulant blood samples were collected and genomic DNA(g DNA was extracted. A series of clinical examinations were performed on the proband (鈪，

本文編號：1884402