Detection of Escherichia coli O157:H7 using chicken immunoglobulin Y.
A sandwich ELISA technique was examined to detect Escherichia coli O157:H7 using chicken anti-E. coli O157:H7 IgY as the capture-antibody and an anti-E. coli O157 mouse mAb conjugated with biotin as the detection antibody. The anti-E. coli O157:H7 IgY was harvested from eggs laid by hens (23 weeks of age, Single Comb White Leghorn) immunized with formalin-killed E. coli O157:H7. The IgY was purified by water dilution methods and gel chromatography on Sephacryl S-300 followed by ammonium sulfate precipitation. The sensitivity (CFU/ml) of sandwich ELISA for the E. coli O157:H7 was repeatedly examined with 10 replicates of each sample and a standard curve was plotted. The sandwich ELISA can detect as low as 40CFU/ml of E. coli O157:H7. The data suggest that chicken IgY-based sandwich ELISA provides a reliable, inexpensive and sensitive assay for the detection of the food-borne pathogen E. coli O157:H7.
Sunwoo
Hoon H
HH
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Canada AB T6G 2P5. hsunwoo@ualberta.ca <hsunwoo@ualberta.ca>
Wang
Welson W
WW
Sim
Jeong S
JS
D000818:Animals; D000907:Antibodies, Bacterial / Q000737:chemistry* / Q000276:immunology; D015373:Bacterial Typing Techniques / Q000379:methods; D002645:Chickens / Q000276:immunology / Q000382:microbiology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D019453:Escherichia coli O157 / Q000737:chemistry* / Q000276:immunology; D005516:Food Microbiology; D007136:Immunoglobulins / Q000737:chemistry* / Q000276:immunology; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; C016512:IgY; D007136:Immunoglobulins