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研究生:張祐泟
研究生(外文):You-Cheng Chang
論文名稱:鳥型複合分枝桿菌的基因分型與肺部疾病的相關性
論文名稱(外文):Association between strain genotyping and lung disease of Mycobacterium avium complex
指導教授:許鴻猷
指導教授(外文):Hung-Yu Shu
學位類別:碩士
校院名稱:長榮大學
系所名稱:生物科技學系(所)
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:中文
論文頁數:60
中文關鍵詞:鳥型複合分枝桿菌
外文關鍵詞:Mycobacterium avium complex
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非結核分枝桿菌在全世界所造成的感染快速地增加中,在台灣以鳥型複合分枝桿菌被分離出來的比率最高。鳥型複合分枝桿菌會導致淋巴炎、肺部疾病和瀰漫性疾病。早期鳥型複合分枝桿菌被分成3群,包括鳥型分枝桿菌、胞內分枝桿菌、和其它(非前兩者)。使用傳統培養的方法或生化檢測都無法將其鳥型複合分枝桿菌加以分型,但以基因型方法可將鳥型複合分枝桿菌分出9種不同菌種,先前研究指出在不同地區的病患的菌株會有所差異。署立胸腔病院收集了184株鳥型複合分枝桿菌,本研究以兩段基因序列:internal transcribed spacer regions between the 16S and 23S rRNA genes (16S-23S ITS) 和 rpoB gene 作為菌株分型的依據。在184株菌株,我們成功定序了181株rpoB gene,而以16S-23S ITS 分析共成功定序176株,由定序分析的結果得知胞內分枝桿菌為感染肺部疾病中佔最多數。先前研究顯示鳥型複合分枝桿菌所致疾病中以鳥型分枝桿菌鳥型亞種和胞內分枝桿菌為主,而在我們的研究結果中發現在南台灣以胞內分枝桿菌佔最多數。

Nontuberculous mycobacteria (NTM) infections have increased rapidly throughout the world. One type of NTM infection caused by Mycobacterium avium complex (MAC) has been frequently isolated in Taiwan. This opportunistic human pathogenic MAC causes lymphadenitis, pulmonary disease, and disseminated disease. Previously, MAC species were classified into three main subgroups (M. avium, M. intracellulare, and the others), but their phenotypes were indistinguishable when conventional culturation methods and biochemical tests were used. Recently, MAC has found to have nine distinct species using genetic screening. The isolates of MAC species differ between patients and have a correlation with their geographical locations. We have collected 184 MAC isolates from the Chest Hospital, a hospital under control of the government of Taiwan’s Department of Health. To differentiate these isolates, the internal transcribed spacer regions between the 16S and 23S rRNA genes (16S-23S ITS) and the rpoB gene have been amplified and sequenced. Phylogenetic analysis based on 16S-23S ITS and rpoB gene sequences were also performed. Out of the 184 MAC isolates, 181 isolates’ rpoB gene were identified and 176 isolates’ 16S-23S ITS were determined. Based on the findings from the 16S-23S ITS sequences, M. intracellulare were the most frequently isolated types and were the most common cause of pulmonary disease. Several studies have revealed that M. avium subsp. avium and M. intracellulare are the most frequent isolates from patients with pulmonary infections among MAC species worldwide. Our results revealed that M. intracellulare was the most frequently isolated type in patients with pulmonary infections among MAC isolated from southern Taiwan.

壹、緒論 1
貳、文獻探討 5
2-1 分枝桿菌和疾病之探討 5
2-2 影像學檢查對於非結核性分枝桿菌所致肺部疾病之診斷 7
2-3 非結核性分枝桿菌菌株的分類 9
?、研究方法 12
3-1實驗流程 12
3-2實驗材料 12
3-3實驗方法 13
3-3-1 收集鳥型複合分枝桿菌檢體 13
3-3-2 再培養鳥型複合分枝桿菌 13
3-3-3 取出菌體 14
3-3-4 滅菌之處理及抽取去氧核醣核酸 14
3-3-5 設計出聚合酵素鏈鎖反應所要用的兩對引子 15
3-3-6 聚合酵素鏈鎖反應(Polymerase chain reaction)的方法及條件 16
3-3-7 以電泳膠確認已得到聚合酵素鏈鎖反應的產物 16
3-3-8 將聚合酵素鏈鎖反應的產物送至陽明大學基因體中心作定序分析 16
3-3-9 檢查序列分析圖 17
3-3-10 將正向序列和反向序列組合 17
3-4 病歷整理病人相關的資料 17
3-5 以演化樹來分族群 17
3-6 統計方法計算各分群的疾病勝算比 17
肆、研究結果: 19
4-1 PCR 產物的分析 19
4-2 有無肺部疾病的分類 21
4-3 以演化樹作分群 23
伍、個案報告 36
陸、討論 44
6-1討論 44
6-1-1分枝桿菌的特性及其重要性 44
6-1-2 分枝桿菌和疾病的相關性 45
6-1-3 鳥型複合分枝桿菌分型的研究 46
6-1-4 影像學對於鳥型複合分枝桿菌所致肺部疾病的診斷 47
6-1-5 鳥型複合分枝桿菌的分群和肺部疾病的相關性 48
6-2 未來展望 49
參考文獻 50
附錄 56


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