Grow E. coli 0157:H7 in trypticase soy broth at 37°C for 18 h; add 0.5% formalin and keep at 37°C for 2 weeks. Inoculate formalized culture into rabbits through ear vein. Give a total of 5 inoculations (0.5, 1.0, 2.0, 4.0 and 4.0 ml), one each, at 5-day intervals. Remove blood after 6 weeks and obtain serum. Heat serum at 56°C for 30 min and adsorb antiserum 6 times with about 1012 cells of heat-treated (121°C, 1 h) E. coli 0157:H7 (1 ml packed cells to 20 ml antiserum). Use antibody solution at 1:5000 dilution.
Original Source: Bacteriological Analytical Manual, 8th Edition, Revision A, 1998.