大肠杆菌O157:H7抗体胶体金免疫层析检测试纸条的研制

Development of a Immunochromatographic Strip for the detection of antibodies against enterohemorrhagic E.coli O157:H7 in mice

为建立快速检测鼠感染大肠杆菌O157∶H7后的抗体,本研究采用颗粒为20nm的胶体金溶液标记纯化的羊抗鼠IgG,制备金标探针,喷涂于玻璃纤维膜上;将大肠杆菌O157∶H7抗原和抗大肠杆菌O157∶H7单克隆抗体F1分别包被于硝酸纤维素膜上作为检测线和质控线,组装成鼠血清O157∶H7抗体胶体金免疫层析检测试纸条。实验结果表明,该试纸条仅与O157∶H7阳性鼠血清发生特异性反应,用该试纸条与常规间接ELISA平行比较检测O157∶H7免疫鼠血清,抗体效价分别为1×106和1×105,两者的敏感度相当。该试纸条操作简便,10min可出结果,适用于调查与追溯鼠O157∶H7感染状况时的现场检测。

To develop a rapid and accurate test based on the immunochromatographic（IC） assay for the detection of antibodies against enterohemorrhagic E.coli O157 ∶H7 in mice,the probes were prepared by coupling 20 nm gold particles in colloidal solution with purified goat anti-mouse IgG,and dispensed on the glass fibers.The EHEC O157∶H7 antigen and mouse anti-EHEC O157∶H7 monoclonal antibody F1 were separately coated on the nitrocellulose membrane as the test line（T line） and the control line（C line）.The rapid IC strip was assembled in regular sequence.Tests on serum samples showed that only O157∶H7 positive serum produced a positive signal on the IC strip.The sensitivity of the strips was similar to the indirect ELISA.The IC strip developed in this study is specific,simple and sensitive,and suitable for the field investigation of EHEC O157∶H7 infection in mice.