A MICROTRAY‐BASED AGGREGATION ASSAY FOR THE RAPID DETECTION OF POLYMERASE CHAIN REACTION AMPLICONS PRODUCED FROM BACTERIAL PATHOGENS

Abstract

ABSTRACT 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.