- 學位論文
建立裘馨氏肌肉萎縮症和貝克氏肌肉萎縮症之全方位基因分析策略
Comprehensive Genetic Analysis in Duchenne/Becker Muscular Dystrophy
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摘要
裘馨氏肌肉萎縮症(Duchenne muscular dystrophy)和貝克氏肌肉萎縮症(Becker muscular dystrophy)，是一種性聯隱性遺傳病，也是兒童期最常見的肌肉萎縮症。致病原因是由於dystrophin基因突變導致肌肉細胞不能正常產生肌縮蛋白，會使鈣離子滲入細胞，使肌纖維壞死，進而導致患者全身肌肉無力。 結合臨床診斷和基因檢測以利裘馨氏肌肉萎縮症或貝克氏肌肉萎縮症準確且有效率診斷流程。建立台灣裘馨氏肌肉萎縮症和貝克氏肌肉萎縮症之資料庫，了解台灣地區是否有特定突變區域，已利提供未來治療的方針。因dystrophin基因非常龐大，單點突變的偵測非常費時，以高分辨熔解分析技術（High-Resolution Melting, HRM）針對dystrophin基因進行分析，可以減短基因檢測分析時間並提供準確基因分析結果。 有288家庭總共551個案利用Multiplex Ligation-dependent Probe Amplification , MLPA和multiplex PCR進行血液基因檢測，結果為突變模式是大片段外顯子的缺失有99的家庭數，大片段外顯子的重覆有39的家庭數。利用以高分辨熔解分析技術（High-Resolution Melting；HRM）和Single-condition amplification/internal primer (SCAIP )進行基因檢測，有72的家庭數是小片段缺失或重複和點突變。 因此，結合臨床診斷和全方位的基因檢測流程能提高裘馨氏肌肉萎縮症或貝克氏肌肉萎縮症的診斷率；根據目前結果能初步建立台灣之裘馨氏肌肉萎縮症或貝克氏肌肉萎縮症的資料庫，希望進而能運用於未來治療或幫助裘馨氏肌肉萎縮症與貝克氏肌肉萎縮症的家庭進行遺傳諮詢。
並列摘要
Duchenne muscular dystrophy, DMD and Becker muscular dystrophy, BMD are severe recessive X-link muscular dystrophies that commonly occur during childhood. The disorder is caused by a mutation of the dystrophin gene (DMD gene) which encoded the dystrophin. Absence of dystrophin leads to excess calcium to penetrate the sarcolemma, which is called the cascading process, and results in muscle weakness overall. In order to provide a more accurate and efficient procedure of diagnosis, we shall combine clinical diagnosis with genetic examination. By establishing the DMD and BMD database among Taiwanese, we can identify the hotspots mutation, and thus provide potential treatments. The detection of point mutations is quite time-consuming because of the huge size of DMD gene. By the application of High Resolution Melting (HRM) analysis, we can reduce the time for examination and also provide accurate results. With Multiplex Ligation-dependent Probe Amplification (MLPA) and multiplex PCR, we examined 288 families, 551 individual, for level I genetic analysis. Deletions of one or more exons account for 99 of families, and duplication of one or more exons account for 39 of families. Furthermore, we apply HRM analysis and Single-condition amplification/internal primer (SCAIP) to examine the level II genetic analysis, and observed 72 families with exonic small deletion, duplication and point mutation. The diagnostic yield of DMD and BMD can be improved with cooperating comprehensive genetic approaches and clinical symptom/signs. We have established the genetic database of DMD/BMD in Taiwan, which may help the therapeutic potential and genetic counseling in the families with DMD/BMD.
參考文獻
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