检测产肠毒素性大肠杆菌粘附素抗体间接ELISA的建立

Development of an indirect ELISA for detecting antibodies to adhesins of ETEC

为了评价仔猪大肠杆菌病K88-K99-987P-F41 四价亚单位疫苗的免疫效果,利用纯化粘附素为包被抗原,建立了检测产肠毒素性大肠杆菌粘附素抗体的间接ELISA。初步确定了各种反应条件: K88、K99、987P、F41粘附素抗原最适包被浓度依次为1∶400、1∶80、1∶40、1∶40,相应粘附素抗体最佳稀释度依次为1∶400、1∶200、1∶200、1∶200。经交叉试验、阻断试验和重复试验证实,本方法具有良好的特异性和重复性。

An indirect ELISA for detecting antibodies to an K88-K99-987P-F41 subunit vaccine against piglet colibacillosis was established by using purified adhesions for coating antigens. Conditions of reaction were as follows: The optimal coating concentration of K88, K99, 987P and F41 antigens was 1∶400,1∶80,1∶40 and 1∶40, respectively.The optimal dilution of corresponding serum samples was 1∶400, 1∶200, 1∶200 and 1∶200, respectively. It was proved that the method was rather specific and repeatable by cross experiment, blocking test and repeated experiment.