本文選題：遺傳性痙攣性截癱 + 全外顯子測(cè)序　； 參考：《青島大學(xué)》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(jìn)行性肌肉緊張、肌無(wú)力和痙攣狀態(tài)。本研究旨在應(yīng)用全外顯子測(cè)序?qū)σ粋�(gè)中國(guó)漢族HSP家系的致病基因進(jìn)行研究,為家系患者做出準(zhǔn)確的基因診斷,為產(chǎn)前診斷和HSP致病機(jī)理的研究奠定基礎(chǔ)。方法收集一個(gè)包含5代32人的HSP家系,繪制標(biāo)準(zhǔn)家系圖,選取該家系Ⅲ代7人、Ⅳ代12人作為研究對(duì)象,采集外周抗凝血樣并提取基因組DNA(g DNA)。對(duì)先證者(Ⅳ11)、先證者患病姐姐(Ⅳ8)和先證者未患病哥哥(Ⅳ9)進(jìn)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。選取Ⅳ11、Ⅳ8和Ⅳ9的g DNA樣本進(jìn)行全外顯子測(cè)序,運(yùn)用SIFT、Poly Phen2、DNAMAN等生物信息學(xué)分析軟件獲取可疑致病基因突變位點(diǎn)。應(yīng)用PCR擴(kuò)增及Sanger測(cè)序的方法將可疑致病突變位點(diǎn)在該家系19名研究對(duì)象和50例與該家系無(wú)血緣關(guān)系的健康對(duì)照者中進(jìn)行驗(yàn)證,分析該位點(diǎn)在家系內(nèi)是否呈共分離,從而進(jìn)一步驗(yàn)證該位點(diǎn)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(jìn)行性痙攣性截癱,無(wú)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(jiàn)明顯異常�；驒z測(cè)結(jié)果:通過(guò)全外顯子測(cè)序和Sanger測(cè)序驗(yàn)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(diǎn)(c.1606CT,p.Gln536X),家系中13例正常者和50例與該家系無(wú)血緣關(guān)系的健康對(duì)照者中均未發(fā)現(xiàn)該突變,表明這一突變位點(diǎn)在該家系中呈共分離。此外,該突變位點(diǎn)在國(guó)內(nèi)外尚未報(bào)道,是一個(gè)新發(fā)現(xiàn)突變位點(diǎn)。結(jié)論結(jié)合臨床癥狀和基因檢測(cè)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(jī)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，

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