Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/49159
標題: 以淋巴增殖反應檢測牛隻結核病之評估試驗 (II)
Development of Lymphocyte Immunostimulation Assay for Detecting Bovine Tuberculosis (II)
作者: 李維誠
關鍵字: 技術發展
畜牧獸醫類
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)。牛型分枝桿菌可藉由家畜傳排泄物及生乳染給人及其他動物;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,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,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免疫反應主要藉由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。因此結核病之感染,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,已不再適用於今日。近年來新研發之牛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,因此較為客觀且可隨時進行複檢,但由於檢驗套組所需之費用甚高,不適合大量樣品之篩檢。因此本計劃在評估以全血模擬化之淋巴球免疫剌激試驗,配合96孔細胞收集器及microbetacounter系統來檢驗牛結核病之可行性,以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為主,因此可藉由,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 within 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 activation of memory T cells but many factors by PPD and then cascade to activate macrophage producing TNF-a and causing skin swelling. This may be interfered by some factors and causes the fause positive and negative results for diagnosis in animals. Interferon-g (IFN-g) 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-g kit 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 modified in whole blood to detect TB infection and compare the results with IFN-g ELISA test and intradermal 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/11455/49159
其他識別: 91農科-7.2.1-檢-B2(6)
文章連結: http://grbsearch.stpi.narl.org.tw/GRB/result.jsp?id=935284&plan_no=91%E8%BE%B2%E7%A7%91-7.2.1-%E6%AA%A2-B2%286%29&plan_year=91&projkey=PG9306-1671&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+%28II%29
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