A microtray-based aggregation assay for the rapid detection of polymerase chain reaction amplicons produced from bacterial pathogens

A microtray-based aggregation test was developed for the rapid and cost-effective detection of polymerase chain reaction (PCR) amplicons from foodborne pathogens. The PCR was performed using biotinylated primers, and the amplified DNA fragments were able to aggregate streptavidin-coated particles. When transferred to a microtyping tray, the aggregated particles converged at the periphery of the well floor to form a visible ring within 5 min. No additional equipment was needed, and purification of the PCR amplicons was unnecessary. Using species-specific biotinylated primers, we successfully demonstrated that the aggregation test can detect and identify Listeria monocytogenes, Salmonella Typhimurium, Staphylococcus aureus, Campylobacter jejuni and Escherichia coli O157:H7 cells. The simplicity rapidity and economy of this method should benefit food-safety monitoring, as well as various other diagnostic PCR experiments.