Detection of <i>Salmonella enteritidis</i> and <i>Salmonella typhimurium</i> in foods using a rapid, multiplex real‐time recombinase polymerase amplification assay
Junan Ren, Yan Man, An Li, Gang Liang, Xinxin Jin, Ligang Pan
Abstract
Abstract Salmonella has been recognized as a major foodborne pathogen for humans and animals. In this study, a multiplex real‐time recombinase polymerase amplification (RPA) was developed for simultaneous detection of Salmonella enterica serovars, Salmonella enteritidis and Salmonella typhimurium , from chicken, eggs, lettuce, and papaya. The reaction was performed for 20 min at 35°C, and the detection limit of the assay was 10 2 CFU/ml for pure culture. In food application, the limit of detection (LOD) of S. enteritidis and S . typhimurium using multiplex real‐time RPA without enrichment procedure was 10 2 CFU/25 g, respectively. After enrichment, the LOD of S. enteritidis and S . typhimurium was 10 CFU/25 g. Moreover, the result for Salmonella spp. was not significantly different from those obtained using a culture‐based method. Additionally, the assay has a lower cross‐reactivity with other pathogenic microorganisms and a good stability performance. Thus, the developed multiplex RPA assay could be used as a rapid tool for the detection of S. enteritidis and S. typhimurium in food.