One-step immunoassay based on filtration for detection of food poisoning-related bacteria

Abstract

A one-step immunoassay based on filtration was presented, which used microbeads for target analyte detection and filters with appropriate pore sizes to distinguish the complexity of target analyte and microbeads. For effective bacterial detection, the microbead size and the filter's pore size must be optimized. The optimal con-centrations of the enzyme (urease) and antibody were determined at the maximum absorbance change, that is, the maximum pH change. The pH change was measured using a field-effect transistor (FET). The correlation between pH change and threshold voltage was estimated to be 21.7 mV/pH, and the correlation between pH change and the source-drain current was estimated to be-379 nA/pH. For the one-step immunoassay, antibodies against target bacteria were isolated from horse serum by filtration, and these antibodies were estimated to have a sufficiently high specificity to overcome cross-reactivity among five types of food poisoning-related bacteria: Escherichia coli O157, Salmonella typhimurium, Listeria monocytogenes, Bacillus cereus, and Staphylococcus aureus. Finally, the FET-based one-step immunoassay was demonstrated for five types of food poisoning-related bacteria in human serum.