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研究生:趙芳瑜
研究生(外文):Fang-Yu Chao
論文名稱:台灣地區幽門螺旋桿菌臨床菌株之血型抗原結合附著因子基因型與臨床疾病的關係
論文名稱(外文):Clinical relevance of the blood-group antigen-binding adhesin genotype of Helicobacter pylori in Taiwanese clinical isolates
指導教授:王雯靜
學位類別:碩士
校院名稱:國立清華大學
系所名稱:生命科學系
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:73
中文關鍵詞:幽門螺旋桿菌血型抗原結合附著因子
外文關鍵詞:Helicobacter pyloriblood-group antigen-binding adhesin
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幽門螺旋桿菌是一種螺旋狀微好氧的革蘭氏陰性菌,寄生於人體的胃表皮黏膜上,已被發現與許多消化道疾病的發生有關,如胃炎、胃潰瘍、十二指腸潰瘍、胃癌。幽門螺旋桿菌可以在細胞膜上表現血型抗原結合附著因子(blood-group antigen binding adhesin,BabA),提供幽門螺旋桿菌附著在宿主細胞上的附著力。而幽門螺旋桿菌特有的液胞毒素(vacuolating toxin)則可以使得細胞發生腫脹,進而脹破死亡。本實驗室之前曾針對台灣地區分離菌株之bab基因型作了初步分析,本論文更進一步延伸bab基因型的分析工作,藉以了解台灣地區臨床菌株的致病因子bab、vacA中間區域基因型和不同消化道疾病的關係。結果發現babA的普行率為73.23%(n=127),且不同臨床疾病群中沒有顯著差異存在。而來自胃癌的幽門螺旋桿菌菌株中,屬於vacAm1和vacAm1/babA+基因型的比例和來自十二指腸潰瘍的菌株有顯著差異(p值分別為0.011和0.025),顯示vacAm1和vacAm1/babA+基因型或許可作為評估罹患胃癌的指標。所有的臨床菌株中,屬於vacAm1/babA+的菌株有43.75%來自胃癌患者,顯示vacAm1/babA+基因型或許可以用來評估罹患消化道疾病患者的患病嚴重程度。此外,在大腸桿菌中表現菌株v344的BabA中間區域,並純化重組蛋白質以製作多株抗體及單株抗體。得到的多株抗體可以辨識為babA+基因型菌株中的BabA蛋白質,而現在也已成功地將融合瘤細胞株2G2和16C4注射到小鼠中以取得單株抗體,將有助於進一步了解幽門螺旋桿菌附著到上皮細胞的作用。

Helicobacter pylori is a spiral microaerophilic Gram-negative bacterium which colonizes the mucus layer overlaying the gastric epithelium. Infection with H. pylori is associated with several gastrointestinal diseases, including gastritis, gastric ulcer, duodenal ulcer and gastric cancer. The blood-group antigen-binding adhesin (BabA) targeting human Lewisb surface epitopes is considered as an essential factor to mediate the epithelial attachment of H. pylori. Vacuolating toxin (VacA) is a protein of 87 kDa that has been shown to vacuolated epithelial cells. The aim of this study was to investigate the relationship between bab genotypes of H. pylori and different gastrointestinal diseases and to generate antibodies against BabA for further biochemical studies. The relationship with another clinically related cytotoxin, VacA, was also studied. The babA gene was detected in 73.23% of 127 clinical isolates. No significant difference was seen among different diseases. The proportion of vacAm1 (p = 0.011) and vacAm1/babA+ (p = 0.025) showed significant difference from gastric cancer and duodenal ulcer, respectively suggesting vacAm1 and vacAm1/babA+ genotype might be useful as a marker for gastric cancer. An intein-fused protein of BabA middle region from strain v344 (344L) was expressed in Escherichia coli. Polyclonal antibody raised against 344L was able to detect BabA in the crude extracts of babA+ strains. Two monoclonal antibodies against 344L, 2G2 and 16C4, were also obtained which would be useful to study the mechanism of H. pylori adherence to gastric epithelium.

中文摘……………………………………………………………………………i
英文摘……………………………………………………………………………ii
壹、前言
一、幽門螺旋桿菌的發現……………………………………………………1
二、幽門螺旋桿菌的流行病學意義…………………………………………1
三、幽門螺旋桿菌與消化道疾病的相關性…………………………………2
四、幽門螺旋桿菌的致病因子………………………………………………4
五、血型抗原結合附著因子(Blood-group antigen binding adhesin, BabA) …………………………………………………………………………6
六、bab核苷酸序列之分析與比對…………………………………7
七、研究目標…………………………………………………………………8
貳、材料與方法………………………………………………………………11
參、結果
一、台灣地區幽門螺旋桿菌bab基因之比較與分析……………………… 15
二、分析台灣地區幽門螺旋桿菌bab中間區域基因型…………………… 16
三、幽門螺旋桿菌之bab、vacA中間區域基因型與不同消化道疾病的相關性………………………………………………………………………………18
四、從罹患肝方面疾病病患所取得的幽門螺旋桿菌vacA、bab基因型與來自於消化道患者的菌株之差異………………………………………………22
五、肝硬化病人受幽門螺旋桿菌多重感染的情形…………………………23
六、生產可辨識Bab蛋白質的多株抗體及單株抗體……………………… 24
肆、討論………………………………………………………………………27
伍、參考文……………………………………………………………………33
陸、圖表………………………………………………………………………39

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