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Real-Time Label-Free Acoustic Technology for Rapid Detection of Escherichia coli O157:H7

(Akubio Limited, Cambridge, UK;

^aaddress correspondence to this author at : Akubio Limited, 181 Cambridge Science Park, Cambridge, CB4 0GJ, UK; fax 44-1223-225336, e-mail lhuang@akubio.com)

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Escherichia coli O157:H7 (O157) is a rare serotype of E. coli that produces large quantities of a powerful toxin that can severely damage the lining of the intestine and induce extreme diarrhea, hemorrhagic colitis, and kidney damage. People are commonly infected by food or water; because bacteria can live asymptomatically in healthy ruminant mammals, meats can become contaminated during processing (1)(2). A rapid test to identify contaminated foodstuffs, water supplies, and infected persons would assist in reducing the risk of spreading this disease.

Many assays have been developed for detection of O157 (3)(4)(5)(6). These methods are time-consuming, however, if they require bacterial culture and may be too complex and costly for use in routine analysis. Rapid confirmation PCR assays for O157 have recently been developed (7)(8), but inhibition of PCR by humic compounds present in complex matrices such as food and surface waters can lead to difficulties such as false-negative results. To address these shortcomings, we developed a rapid, specific proof-of-principle assay for the detection of E. coli O157:H7 with Resonant Acoustic Profiling^TM (RAP) technology (9).

We obtained heat-inactivated E. coli O157:H7 cells from Kirkegaard and Perry Laboratory. Five clinical E. coli isolates provided by Dr Derek Brown (Addenbrookes, Cambridge, UK) and an American Type Culture Collection E. coli 25922 were used as negative controls. Five of these isolates were live bacteria, and heat-inactivated E. coli 170044 was used as a killed control. Affinity-purified goat anti-O157 polyclonal antibodies (pAbs) were obtained from Kirkegaard and Perry Laboratory. Anti-O157 pAb was isolated from a pooled serum from goats immunized with heat-killed whole cells of E. coli O157:H7. The lyophilized anti-O157 antibody was reconstituted with 0.3 mol/L sodium phosphate buffer (0.1 mol/L NaH₂PO₄, . . . [Full Text of this Article]