生物素——亲和素酶联免疫吸附测定法检测水稻白叶枯病病菌

DETECTION OF XANTHOMONAS CAMPESTRIS PV. ORYZAE USING ABC-McAb-ELISA

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。

A technique, ABC-McAb-ELISA for detection of Xanthomonas campestris pv. oryzae was developed by adding the biotin-avidin system(BAS) to the enzyme linked immunosorbent assay(ELISA) which applied the monoclonal antibodies(McAbs) for X. campestris pv. oryzae as the antibody or the second antibody. ABC-McAb-ELISA could detect the bacteria in different plant materials effectively. The scnsitivety of ABC-McAb-ELISA (10~3cells/ml) was higher than that of McAb-ELISA (about 10~5cells/ml). Morever, ABC-McAb-ELISA did not change the specifity of McAbs.