Lateral flow assay strip for detection of Escherichia coli O157:H7.

Abstract

The use of polyclonal antibody (IgG) has recently been applied to the detection of bacteria. We developed a lateral flow assay (LFA) strip using a specific IgG in combination with colloidal gold on a nitrocellulose membrane. A conjugate, gold-anti Escherichia coli (E. coli) O157:H7 IgG was developed in this study for the detection of E. coli O157:H7 in food. The 40 nm in size of colloidal gold nanoparticles was used to conjugate the anti-E. coli O157:H7 IgG. The optimal concentration, 12.0 μg/ml of the anti-E. coli O157:H7 IgG was determined by standard curve generated in titration method. The serially diluted of E. coli O157:H7 was detected and clearly visualized on the LFA strip as low as 106 CFU/ml (result not shown). The IgG raised in rabbit have shown specific binding capacity against E. coli O157:H7. No other genus of bacteria, including Salmonella typhimurium, Listeria monocytogenes and Campylobacter jejuni reacted to the IgG. The LFA strip could also detect E. coli O157:H7 in different food samples matrices after 18 h-enrichment and this result were in accordance with the results of the polymerase chain reaction (PCR) and colony count.