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以纯化培养的甘蔗宿根矮化病(RSD)病菌作为抗原,免疫健康家兔,制备了RSD多克隆抗体。采用间接酶联免疫吸附试验检测其效价,并通过斑点酶标免疫法(Dot blot enzyme immunoassays,DB-EIA)确定其工作浓度,同时与澳大利亚的抗体比较。结果表明,制备的RSD多克隆抗体效价大于1∶10000,在用DB-EIA法检测样本时的工作浓度以稀释10倍较好;与澳大利亚抗体的检测结果一致,但在印迹着色方面比澳大利亚的浅一些。试验结果表明,自制RSD抗体的效价略低于澳大利亚RSD抗体,但检测效果较好,可以满足今后在甘蔗生产上对RSD的检测要求。
The RSD polyclonal antibody was purified from the cultured Saccharomyces cerevisiae, and the rabbits were immunized. The titer was detected by indirect enzyme-linked immunosorbent assay (ELISA), and its working concentration was determined by Dot-blot enzyme immunoassay (DB-EIA). Meanwhile, it was compared with that of Australian antibody. The results showed that the titer of the prepared polyclonal antibody against RSD was greater than 1: 10000, and the working concentration when diluted with DB-EIA was 10 times better than that of the Australian antibody. The results were consistent with those from Australian antibody but lower than those from Australia Shallow. The test results showed that the self-made RSD antibody titer slightly lower than the Australian RSD antibody, but the detection effect is better, to meet the RSD detection requirements in the future sugar cane production.