Application of Monoclonal Antibodies in the Detection of Xanthomonas Albilineans, the Sugarcane Leaf Scald Pathogen

本研究乃利用先前針對甘蔗白條病菌Xanthomonas albilineans研製所得之單元抗體，從事病原菌之偵測工作。研究中共選定MAs 1, 2, 3, 5, 7, 9, 24, 34, 35, 36, 37, 38等12株具特異性之融合瘤單元抗體進行各項測定，而偵測之方式則包括間接免疫酵素聯結法(Indirect ELISA），免疫螢光抗體法(Immunofluorescent staining),及點漬染法(dot-ELISA)等三種方法。在以間接免疫酵素聯結法進行測定時，發現以碳酸鹽緩衝液(Carbonate buffer)稀釋病蔗抽出液二至三倍，並在37 C通風狀態下過夜以從事抗原吸附，最能有效地偵測到蔗汁中之病原菌，而抗體中以MAs 1, 3, 9, 34, 35, 36, 38等七株抗體具較佳之偵測能力。若以免疫螢光抗體法進行偵測時，則MAs 1, 2, 3, 5, 34, 37, 38等七株單元抗體能完整的染出整個菌體，而MAs 7, 9, 24, 35, 36等五株單元抗體則僅能在菌體上呈現點狀之螢光染色，較不適宜應用於病原菌之偵測。而以點漬染法從事病原菌偵測時，則僅有MAs 1, 2, 3等三株單元抗體能專一地偵測出病株抽出液中之病原細菌。

關鍵字

病害偵測:單元抗體；甘蔗白條病

並列摘要

Specific monoclonal antibodies against the sugarcane leaf scald pathogen, Xanthomonas albilineans, developed in our previous study were applied in the disease detection. Specific monoclonal antibodies obtained from 12 hybridoma clones, MAs 2, 7, 35, 36, 37 and 38, were used in the serological detection procedures including enzyme-linked immunosorbent assay (ELISA), immunofluorescent staining, and dot-ELISA. For ELISA, when the sugarcane sap extracted from diseased canes was diluted two- to three-fold with 0.05 M carbonate buffer (pH 9.6) and then coated under 37 C along with overnight ventilation, the pathogen in the sap can be readily detected by MAs 1, 3, 9, 34, 35, 36 and 38. For immunofluorescent staining, MAs 1, 2, 3, 5, 34, 37 and 38 can detect X. albilineans both in the cultured X. albilineans preparation and the diseased sugarcane sap effectively, while MAs 7, 9, 24, 35 and 36 can only stain the cultured bacteria sparsely and weakly. When dot-ELISA was applied, only MAs 1, 2 and 3 can detect the X. albilineans present in the sugarcane sap from diseased canes.

並列關鍵字

disease detection ； monoclonal antibodies ； sugarcane leaf scald ； Xanthomonas albilineans

國際替代計量

Application of Monoclonal Antibodies in the Detection of Xanthomonas Albilineans, the Sugarcane Leaf Scald Pathogen

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