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Detection of Escherichia coli O157:H7 using chicken immunoglobulin Y.

Posted by on in 2006
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Sunwoo HH, Wang WW, Sim JS.
Immunol Lett. 2006 Aug 15;106(2):191-3. Epub 2006 Jun 2.
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Canada AB T6G 2P5. hsunwoo@ualberta.ca <hsunwoo@ualberta.ca>


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.

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