POLYMACRON(TM) ENZYME-IMMUNOASSAY SYSTEM FOR DETECTION OF ESCHERICHIA-COLI O157 INOCULATED INTO FOODS

A rapid and simple enzyme immunoassay system was developed for detecti on of the food-borne pathogen Escherichia coli O157. This system is ba sed on the use of anti-E. coli O157 antibody-coated Polymacron(TM), an inexpensive macroporous polyester fabric, as a high-surface-area immu noadsorbent for the rapid capture and subsequent immunoenzymatic detec tion of E. coli O157 antigens extracted from test samples by heating a t 100 degrees C for 10 min in the presence of sodium cholate. A dot bl ot format was used which facilitated the assay of multiple samples. Th e method was specific for all strains tested bearing the O157 and rela ted antigens (e.g., group N Salmonella), giving positive reactions in the assay of pure cultures of 29 E. coli O157:H7 strains, one E. coli O157:H12, one E. coli O157:NM (nonmotile) and one Salmonella urbana st rain, but not in the assay of a variety of other gram-negative and gra m-positive bacteria. The method permitted the detection of ca. 400 E. coli O157:H7 CFU in a 5-mu l spot applied to Polymacron(TM), and of as few as 0.4 CFU of E. coli O157.H7 cells per g inoculated into ground beef samples, which were assayed after overnight enrichment.