PRODUCTION AND CHARACTERIZATION OF MONOCLONAL-ANTIBODIES SPECIFIC FORTHE LIPOPOLYSACCHARIDE OF ESCHERICHIA-COLI O157

Identification of the O157 antigen is an essential part of the detecti on of Escherichia coli O157:H7, which is recognized as a major etiolog ic agent of hemorrhagic colitis, However, polyclonal antibodies produc ed against E. coli O157:H7 lipopolysaccharide (LPS) may react,vith sev eral other bacteria including Brucella abortus, Brucella melitensis, Y ersinin enterocolitica O9, Escherichia hermannii, and Stenotrophomonas maltophilia. We produced eight monoclonal antibodies (MAbs) specific for the LPS of E. coli O157, Western blots (immunoblots) of both the p henol phase (smooth) and the aqueous phase (rough) of hot phenol-water -purified LPS indicated that three of the MAbs were specific for the O antigen and five were reactive with the LPS core, The eight MAbs coul d be further differentiated by their reactivities to Salmonella O30 LP S (group N), which is reported to be identical to the E. coli O157 ant igen, All eight MAbs reacted strongly to all of the 64 strains of E, c oli O157 tested, which included 47 isolates of O157:H7 and 17 other O1 57 strains, None of the eight MAbs cross-reacted with any of the 38 ot her E. coli serotypes tested, which consisted of 29 different O-antige n serotypes, or with 38 strains (22 genera) of non-E. coli gram-negati ve enteric bacteria.