Establishment of a method for the simultaneous detection of four foodborne pathogens using high‐throughput suspension array xTAG technology

Abstract

Summary The purpose of this study is to establish a rapid and accurate method based on multiplex polymerase chain reaction ( PCR ) combined with suspension array xTAG (Flexible Sequence‐tagged, xTAG ) technology to simultaneously detect Salmonella Typhimurium, Brucella spp., Bacillus cereus and Shigella spp. in raw milk. The primers were designed based on the specific target gene of the four pathogenic bacteria, and then the TAG sequence was used to modify the primers. First, the products of PCR were hybridised with microspheres labelled with anti‐ TAG , and then they were conjugated to streptavidin phycoerythrin. Finally, the products were detected on a Luminex 200 analyser. The results showed that the detection of milk samples demonstrated 100% specificity. The hybridisation conditions were successfully optimised. We successfully constructed a rapid, simple and high‐throughput method to detect four foodborne pathogens. In conclusion, the established xTAG detection method provides a highly specific and reliable way to detect four kinds of foodborne pathogens in milk products.