水环境中大肠杆菌多特征抗原及抗体的制备研究

为了建立用于水环境中大肠杆菌的酶联免疫快速检测方法,针对水中大肠杆菌属多种血清型的特征,制备了水环境样品中大肠杆菌的多特征抗原,包括全菌体抗原、破碎全菌体抗原、菌体抗原、鞭毛抗原和菌毛抗原;采用这5种大肠杆菌抗原分别免疫新西兰大白兔,获得了5种大肠杆菌多克隆抗体,抗体效价高、纯度好,具有较强且稳定的特异性结合抗原的功能;间接ELISA方法表明,全菌体抗体和破碎全菌体抗体的效价均大于1×10⁵,性能优良.基于上述抗体,建立了水中大肠杆菌间接ELISA检测方法,实际水样的检测结果表明,检测限可达10⁴个/L.

Preparation of E. coli Antigens with Multi-characters and Antibodies in Water Environment

In order to establish enzyme-linked immunosorbent assay(ELISA) method to detect E.coli in water environment, E.coli multi-characters antigens in water environment were prepared according to the characters of kinds of E.coli serotypes, including antigen of whole cell, antigen of disrupted whole cell, somatic antigen, flagellar antigen and fimbrial antigen. E.coli polyclonal antibodies were obtained from the New Zealand rabbits immunized with these five antigens, respectively. Antibodies generated in this research are with high titers and good purity, can conjugate with antigens, specifically, stably and strongly. Indirect ELISA shows the titers of antibody of whole cell and antibody of disrupted whole cell are both more than 1×10⁵, and the antibodies are of good quality. Based on these antibodies, we established indirect ELISA method to detect E.coli in water environment. The result shows that the detection limitation could be 10⁴/L.