Antibodies among healthy population of developing countries against enterohaemorrhagic Escherichia coli O157:H7.

In Thailand, no reports are available on Escherichia coli serotype O157:H7, a causative agent of severe bloody diarrhoea, sometimes associated with haemolytic-uraemic syndrome and thrombotic thrombocytopenic purpura. The reason for the non-identification of infection due to E. coli O157 in this country and in other developing countries has not been rigorously discussed. The aim of this study was to determine the humoral response against the infectious organism. The IgM and IgG antibody responses against E. coli O157 lipopolysaccharide were studied using indirect enzyme-linked immunosorbent assay. Three hundred and thirty-two serum samples obtained from healthy blood donors and patients with diseases unrelated to diarrhoea were investigated. With a cut-off value of mean +2 SDs for each age-group, the frequency of the IgM and IgG responses to O157 lipopolysaccharide was 11.74% (39 of 332 samples) and 22.59% (75 of 332 samples) respectively. Furthermore, agglutination test of 173 subjects revealed titres ranging from 10 to 40 in all the samples. The results suggest possible exposure of the Thai population to cross-reacting antigens from other intestinal organisms in addition to infection due to E. coli O157:H7.

Characterization of recombinant antibodies developed for capturing enterohemorrhagic Escherichia coli O157:H7.

Escherichia coli O157:H7, an emerging cause of food-borne disease with the occurrence of an estimated 20,000 illnesses and 250 deaths each year in the United States, has now been reported from several countries worldwide. Infections with this bacteria, which follows the ingestion of contaminated food by humans, causes bloody diarrhea, hemolytic uremic syndrome (HUS), and renal disease, that can have serious health implications. The source of food contamination is usually associated with animals, mainly cattle. Many cattle become infected early in life when they are exposed to an environment that is contaminated by other animals shedding the organisms in their feces. Detection of E. coli O157:H7 in feces or contaminated food samples requires tests with high sensitivity, which is increased by the use of monoclonal antibodies. However, the production of concentrated monoclonal antibodies in ascites raises animal welfare concerns, and can be expensive. In this study, single chain of variable fragment (scFv) molecules were developed from hybridoma clones that produce immunoglobulins specific for the LPS and flagella antigen of E. coli O157:H7 using phage display technology. The reactivity of the soluble scFv for their respective antigens was preserved in ELISA and by partial inhibition of bacterial agglutination with polyclonal antiserum. Furthermore, the scFv were able to capture E. coli O157:H7 bacteria demonstrating their potential use in diagnostic assays.