樹木褐根病單元抗體檢測技術之建立

樹木褐根病(brown root rot disease)是台灣重要樹木病害之一，由病原真菌Phellinus noxius所引起，其寄主範圍極廣，台灣每年許多珍貴樹種皆因此而枯死，並有逐年擴大趨勢。樹木罹病初期不易以外部病徵診斷，為掌握防治先機與日益增加之診斷與檢測所需，本研究應用融合瘤(hybridoma)技術，開發對褐根病菌具專一性之單元抗體，再利用酵素連結免疫吸附法(enzyme-linked immunosorbent assay=ELISA)，提供一套褐根病快速診斷方法。目前共篩選出14株對褐根病菌反應高，且對健康植物組織與其他相似真菌Ganoderma sp.與Phellinus gilvus反應低之專一性融合瘤細胞株。經單株化由其中選出高力價細胞株（編號P-10^(-5)）生產抗腹水，抗腹水之力價測試可達到10-15。由抗腹水純化之單元抗體應用於田間病樹快速檢測，僅需0.5克根部組織即可檢查有無褐根病菌感染。未來本單元抗體將進一步製成膠金標誌套組(colloidal gold-labeling kit)，以提供更方便、不需儀器，而能立即於田間診斷之樹木褐根病檢測試劑。

關鍵字

褐根病；褐根病菌；單元抗體

並列摘要

Brown root rot caused by a pathogenic fungus, Phellinus noxius, is one of the most severe diseases of forest tree in the world. Many valuable trees are destroyed by this pathogen every year in Taiwan. This disease is not easy to be diagnosed by its incited external symptoms especially in the early infection stage. This study was dedicated to develop monoclonal antibodies against P. noxius with ELISA for rapid and sensitive diagnosis of brown root rot. Totally 14 clones selected from the cell lines of hybridoma showed positive for P. noxius in the ELISA tests whereas they were negative for the other similar species of fungi such as Ganoderma spp. and Phellinus gilvus. Among these 14 cell lines, a clone named ”P-10-5” showing the best sensitivity and specificity was chosen to produce ascetic fluid and its titer can reach to 10-15. The monoclonal antibody derived from P-10-5 was further purified from ascetic fluid and used in the diagnosis of brown root rot. Our results in this paper demonstrated that only 0.5 g of diseased root tissue was enough to obtain the reliable data for the diagnosis in ELISA tests. This monoclonal antibody will be further applied in the preparation of colloidal gold-labeling kit for more convenient diagnosis of brown root rot in the field.

並列關鍵字

Brown root rot disease ； Phellinus noxius ； Monoclonal antibodies

國際替代計量

樹木褐根病單元抗體檢測技術之建立

全文下載

主題瀏覽