Abstract
A membrane separator/bioreactor system was developed for rapid detection of Escherichia coli O157:H7. The system consisted of a membrane separator/bioreactor (0.45 μm of the pore size) to separate the‐complexes of E. coli O157:H7 and alkaline phosphatase‐conjugated anti‐E. coli O157:H7 antibodies from the sample and to produce p‐nitrophenol through the enzymatic reaction (p‐nitrophenyl phosphate hydrolysis), and an optical detector for measuring the p‐nitrophenol absorbance at 400 nm. The membrane material and the flow rate of the substrate for the enzymatic hydrolysis had great effects on the absorbance of p‐nitrophenol. The optimum conditions for the enzymatic reaction were determined as 1.0 M Tris buffer, pH 8.0, and 0.1 M MgCl2 for this system. The detection range was 104± 107 CFU/mL with a relative standard deviation of 4.3 ± 14.2%, and whole procedure could be completed in 50 min without any enrichment and culture. Other bacteria such as Salmonella typhimurium, Campylobacter jejuni and Listeria monocytogenes had no significant interference with the detection of E. coli O157:H7.
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