In order to establish enzyme-linked immunosorbent assay (ELISA) method to detect E. coli in water environment, E. coli multicharacters 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 x 10(5), 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 10(4)/L.