Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/23786
標題: 台灣豬隻E型肝炎病毒分子 之偵測,選殖,與表現
Detection, cloning, and expression of swine HEV in Taiwan
作者: 蔡文瑄
Tsai, Wen-Hsuan
關鍵字: hepatitis E virus
E型肝炎病毒
出版社: 生物化學研究所
摘要: 關於臺灣地區的肝炎研究有報導指出肝炎患者中有不具出國經驗而罹患急性E型肝炎者,由於臺灣地區並非E型肝炎流行疫區,臺灣本地的E型肝炎病毒研究逐漸受重視;為了瞭解動物在E型肝炎病毒宿主上所扮演的角色,家畜如牛、豬等亦成為E型肝炎病毒的研究對象。而研究報導指出,家畜亦發現有E型肝炎病毒的感染,因此本研究將針對臺灣地區飼養較興盛的家畜-豬隻,偵測E型肝炎病毒、選殖其基因並表現選殖之結構蛋白,藉以探討臺灣地區豬隻E型肝炎病毒之感染概況。 本研究以反轉錄聚合酶鏈反應檢測臺灣北、中、南部以及東部等豬場所飼養豬隻之血清或糞便中的E型肝炎病毒;並針對病毒之結構基因選殖出欲表現之結構蛋白基因片段,由具有IPTG誘導特性之載體pRSETc承接此一基因片段,並藉大腸桿菌表現其蛋白。未來並期望可由所產製的結構蛋白發展出用於豬隻E型肝炎感染之 ELISA檢測系統,甚或發展豬隻E型肝炎感染之疫苗。 本實驗共檢測521個豬隻血樣以及54個豬隻糞便檢體,檢測結果發現臺灣地區豬隻感染E型肝炎之比率為1.1~1.7 %;所檢測到的E型肝炎病毒基因型包含type III以及type IV。本實驗並針對type IV之E肝病毒選殖位於其結構基因3’端之部分片段以用於表現蛋白質;其核酸長度約為1.4 kb,蛋白質大小約59 kDa。 本實驗並另外對豬場污水做檢測,發現經靜置發酵處理後之豬場污水並無E型肝炎病毒的陽性反應,顯示有系統的廢水處理有助於防止病毒傳播。由於已有研究報導指出經由糞便及口腔途徑傳染的E型肝炎病毒確實有人畜共通的危險性;因此,在豬隻養殖普遍的臺灣本島有效管理養豬場排水誠係亟待努力的重要課題。而另一方面,在探討本實驗取得檢體的養豬場之養殖系統後,發現感染type III之E型肝炎病毒的仔豬係源自美國進口的種母豬,顯示豬群中E型肝炎的傳播與豬隻貿易之進出口關係密切,對於貿易島國的臺灣而言是不可忽視的疫病管理問題。遺憾的是,目前尚未開發出針對臺灣豬隻E型肝炎病毒感染的免疫檢測系統,因此日後研究當積極研製高效率的抗原蛋白,俾供檢疫及防疫之使用。
Taiwan is not an endemic area of hepatitis E virus (HEV), but there are patients who have not been abroad infected by HEV. It has been reported that farm animals, like pigs and cows, could also be infected by HEV. In order to define the role the farm animals are in being hosts of HEV, this research will focus on pigs, which are the most popular farm animals in Taiwan area. By detection, cloning, and the structural protein expression of HEV, the prevalence of HEV between pigs in Taiwan area would be determined here. In this experiment, samples were sera and stool collected from hog pens in northern, middle, southern, and eastern Taiwan. HEV in the samples was detected by polymerase chain reaction. After cloning the gene of structural protein of HEV into plasmid pRSETc, the protein was expressed by E. coli with IPTG induction. There were 521 sera samples and 54 stool samples detected in this experiment. The HEV infection rate between pigs was found 1.1 % to 1.7 % in Taiwan area. The genotypes of HEV detected in this experiment were type III and type IV. The structural protein expressed in this experiment was 1.4 kb in length and 59 kDa in mass and that was originated from type IV HEV. We also detected the wastewater collected from hog pens, and found that HEV didn't exist in treated wastewater. It reveals that treating waste -water well would help preventing the outbreak of HEV.
URI: http://hdl.handle.net/11455/23786
Appears in Collections:生物化學研究所

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