Novel multiplex PCR assays for rapid identification of Salmonella serogroups B, C1, C2, D, E, S. enteritidis , and S. typhimurium

• Published in: Analytical methods 2022-04, Vol.14 (14), p.1445-1453
• Main Authors: Shang, Yuting, Ye, Qinghua, Wu, Qingping, Xiang, Xinran, Zha, Fei, Du, Mingzhu, Zhang, Jumei
• Format: Article
• Language: English
• Subjects: Assaying; Detection limits; Foodborne diseases; Genes; Genomes; Meat; Multiplex Polymerase Chain Reaction; Multiplexing; Primers; Public health; Pure culture; Salmonella; Salmonella enteritidis - genetics; Serogroup; Target detection
• ISSN: 1759-9660; 1759-9679
• DOI: 10.1039/d1ay02163j

Foodborne illnesses caused by represent a significant public health problem worldwide. The aim of this study was to establish multiplex PCR (mPCR) for the rapid identification of serogroups B, C1, C2, D, and E as well as for the serovars and . Employing pan-genome analysis and PCR verification, , , , , , and four genes ( , , , and ) were identified as specific target genes for serogroups B, C1, C2, D, E, and , respectively. Thereafter, three novel mPCR assays (one of 3-mPCR and two of 2-mPCR) were successfully developed to identify these bacteria based on the target genes and another -specific gene. The primers targeting , , and genes specific to the serogroups C1, C2, and E, respectively, constituted a 3-mPCR, while the other two 2-mPCRs, respectively, consisting primers specific to serogroup D and ( and ), and serogroup B and -specific primers ( and ), were also designed. The specificity of each mPCR was further evaluated by using non-target strains. The detection limits of mPCRs were approximately 10 -10 CFU mL in pure culture and 10 -10 CFU g in spiked chicken meat. In addition, mPCR assays could correctly detect target in food samples. These results suggest that specific targets could be mined efficiently through a pan-genome analysis tool, and the novel mPCR assays developed in this study offer a promising technique for rapid and accurate detection of five serogroups of (B, C1, C2, D, and E) and two serovars ( and ).