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在以单克隆杭体为第2抗体的酶联免疫吸附测定法(ELISA)的基础上,应用生物素——亲和素系统,建立了亲和素——生物素化辣根过氧化物酶复合物——酶联免疫吸附测定法(ABC-ELISA)检测水稻白叶枯病病菌。比较了ABC-ELISA法和常规ELISA法的差异,表明ABC-ELISA法的灵敏度比常规ELISA法高10～100倍。常规ELISA双夹心法的检出量一般为10~6菌体/ml,ABC-ELISA双夹心法达10~菌体/ml;全菌包被ELISA间接法为10~5菌体/ml;而ABC-ELISA间接法为10~3个菌体/ml。ABC-ELISA法并不影响单克隆抗体的高度特异性。用ABC-ELISA法检测58份水稻白叶枯病病叶,16份病种,病叶全部表现阳性反应,病种稻壳如取样在1g以上,检出率较高。
On the basis of enzyme-linked immunosorbent assay (ELISA) with monoclonal antibody as second antibody, avidin-avidin system was used to establish avidin-biotinylated horseradish peroxidase Complex - enzyme-linked immunosorbent assay (ABC-ELISA) for the detection of bacterial blight on rice bacterial blight. The difference between ABC-ELISA and conventional ELISA was compared, indicating that the sensitivity of ABC-ELISA was 10 to 100 times higher than that of conventional ELISA. Conventional ELISA double sandwich method detection volume is generally 10 to 6 cells / ml, ABC-ELISA double sandwich method of 10 ~ cells / ml; whole bacteria coated ELISA indirect method for 10 to 5 cells / ml; and ABC-ELISA indirect method is 10 to 3 cells / ml. ABC-ELISA does not affect the high specificity of monoclonal antibodies. ABC-ELISA method was used to detect 58 bacterial leaf blight of rice disease, 16 disease types, the diseased leaves all showed positive reaction, the detection rate was higher when the disease rice husk was sampled above 1g.