Detection of Escherichia coli O157 in foods by a novel polymyxin-based enzyme-linked immunosorbent assay.
An enzyme-linked immunosorbent assay (ELISA) system was developed using polymyxin immobilized in the wells of a microtiter plate as a high-affinity adsorbent for Escherichia coli O157 lipopolysaccharide (LPS) antigens. Extracts from cell suspensions were reacted with polymyxin-coated microwells followed by immunoenzymatic detection of captured LPS antigens using a commercially available anti-E. coli O157 antibody-peroxidase conjugate and a 3,3',5,5'-tetramethylbenzidine substrate. The polymyxin ELISA was highly sensitive and specific for E. coli strains bearing the O157 antigen. When this ELISA was combined with enrichment, results were in complete agreement with those of standard culture techniques for the detection of this pathogen in a variety of artificially inoculated and naturally contaminated foods. The polymyxin ELISA is a simple and inexpensive assay for E. coli O157 with a 96-well microtiter plate format, making this system ideally suited for processing large numbers of samples.
Blais
Burton W
BW
Canadian Food Inspection Agency, Ottawa Laboratory (Carling), Building 22, Central Experimental Farm, Ottawa, Ontario, Canada. bblais@inspection.gc.ca
Leggate
Johanna
J
Bosley
Jessica
J
Martinez-Perez
Amalia
A
D000907:Antibodies, Bacterial / Q000032:analysis; D000942:Antigens, Bacterial / Q000032:analysis*; D001560:Benzidines; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D019453:Escherichia coli O157 / Q000302:isolation & purification*; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D008070:Lipopolysaccharides / Q000737:chemistry; D011113:Polymyxins; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001560:Benzidines; D008070:Lipopolysaccharides; D011113:Polymyxins; C021758:3,3',5,5'-tetramethylbenzidine