Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/49151
標題: 以淋巴增殖反應檢測牛隻結核病之評估試驗
The Development of Lymphocyte Immunostimulation Assay for Detecting Bovine Tuberculosis
作者: 李維誠
劉正義
簡茂盛
關鍵字: 畜牧獸醫類
應用研究
摘要: 牛結核病是由分枝桿菌屬細菌( Mycobacterium spp. )感染所引起, 在細菌型別上主要分為牛、鳥及人分枝型桿菌( M.bovine, M.avium, M.tuberculosis ).牛型分枝桿菌可由家畜傳染給人及其他動物; 鳥型分枝桿菌則可感染所有品種鳥類、豬與牛; 人型分枝桿菌主要感染人, 但亦可感染豬、牛及狗.牛結核病從古至今, 一直無法被根滅, 並侵害動物及人類健康, 是一種嚴重威脅人畜健康的傳染病, 不但在畜牧事業上造成重大的危害, 且其病原菌可經由空氣與污染之產品、飲水、器械以及其他途徑感染, 尤其是生乳之傳染, 造成嚴重的公共衛生問題.世界各國均將其列為重要之法定傳染病.我國政府自1947年起對台灣地區之乳牛定期實施皮內結核菌素試驗, 對於檢出為陽性反應之牛隻進行撲殺焚燬, 但實施迄今仍無法完全根除本病.在人醫方面, 由於愛滋病流行, 許多愛滋病患因鳥型結核菌或其他非典型菌之感染而使結核病之罹患率再度昇高, 因此迄今結核病仍為相當重要之人畜共通傳染病.結核菌感染宿主之後可存活於非活化狀態的巨噬細胞中, 並在細胞內增殖, 一般的體液性免疫是無法有效清除這類細胞內的病原, 必須藉由細胞免疫反應之活化, 進而激活被感染的細胞本身, 以清除病原.結核菌所引起的細胞媒介性免疫反應呈兩種反應, 一為藉由CD4?T淋巴細胞產生的細胞素, 刺激活化巨噬細胞以殺滅所吞噬的微生物.另一種是藉由CD8?T淋巴細胞執行細胞毒殺作用, 將被感染的細胞溶解.由細胞免疫反應機制, 可知結核病之感染, 可藉由體內是否具有對抗原特異性之記憶性T淋巴細胞之活化及其分泌之產物進行檢測.結核菌素皮內反應試驗法為今日牛結核病最傳統之診斷方法, 但由於其敏感性不高, 且易受多種因素影響, 加上試驗所需時間甚長, 需間隔三個月以上才能再作檢驗, 徒增檢驗之空窗期, 因此已不再適用於今日.近年來澳洲Commonwealth Serum Laboratries公司新發明牛γ-干擾素檢測法, 雖然干擾素試驗是直接採血後, 帶回實驗室操作並藉由機器所判讀, 可避免人為判讀之客觀性, 對可疑病例可重覆檢驗, 因此較為客觀且可縮短檢驗間隔時間, 甚至可隨時進行複檢, 但由於檢驗套組所需之費用甚高, 將造成政府在檢驗經費上之龐大負擔.因此本計畫在於找尋出可替代結核菌素試驗及γ干擾素試驗之檢驗方法, 由於動物感染結核病, 其體內之免液反應是以細胞性免疫應( Cell-mediated immunity, CMI )為主, 因此可藉由淋巴球免疫剌激試驗( Lymphocyte immunostimulation test )來檢驗牛結核病, 其優點為敏感性及特異性均高於結核菌素試驗, 且可在動物體外進行, 較不受動物因子影響, 並可區分鳥型或牛型感染, 縮短檢驗間隔時間及檢驗所需之成本.
The caustive agents of bovine tuberculosis are the Mycobeterium spp.including M.bovine, M.avium, and M.tuberculosis.Mycobeterium spp.are intracellular pathogens which can survival and replication with macrophages to escape the humoral immune response mediated by antibodies.Until now the tuberculosis is also the major health problems in human beings and animals.The cellular mediated immune responses induced by Mycobeterium infection may be through either CD4?T cells to produce cytokines for activating macrophages or through CD8?mediated cytotoxic activity.The memory T cells could also be established after infection that could be utilized for the diagnostic target.The tuberculin skin test is basis on the formation of memory T cells but many factors may interfere and cause the foul positive and negative results for diagnosis in animals.Interferon-γ( IFN-γ )ELISA kit designed by Commonwealth Serum Laboratories for reducing the ambiguous problems.This kit seems to provide a convenient, rapid, and correct processing for diagnosis of Mycobeterium infection; however, the cost of IFN-γkit is extremely expensive to be used as the routine screening.In addition, lymphocyte proliferation method can be used for analysis on cell-mediated immunity.In this study, we try to establish the standard procedures of lymphocyte proliferation analysis after TB infection and compare the results with IFN-γELISA test.The modification of this diagnostic method will also be evaluated for the possibility on massive screening and replace the traditional tuberculin skin test.
URI: http://hdl.handle.net/11455/49151
其他識別: 90農科-6.2.1-檢-BB
文章連結: http://grbsearch.stpi.narl.org.tw/GRB/result.jsp?id=1047238&plan_no=90%E8%BE%B2%E7%A7%91-6.2.1-%E6%AA%A2-BB&plan_year=90&projkey=PG9309-2969&target=plan&highStr=*&check=0&pnchDesc=%E4%BB%A5%E6%B7%8B%E5%B7%B4%E5%A2%9E%E6%AE%96%E5%8F%8D%E6%87%89%E6%AA%A2%E6%B8%AC%E7%89%9B%E9%9A%BB%E7%B5%90%E6%A0%B8%E7%97%85%E4%B9%8B%E8%A9%95%E4%BC%B0%E8%A9%A6%E9%A9%97
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