Abstract
Food-borne pathogens represent a significant threat to global food safety and public health. Enterohemorrhagic Escherichia coli (EHEC) O157:H7, Salmonella, and Cronobacter are among the principal bacterial agents responsible for food-borne illnesses. Advances in sequencing technology have made it possible to identify particular genetic markers for these pathogens through large-scale genomic analysis. In this study, two conserved genes, z0340 and sbcC, were screened in silico from extensive bacterial genomes (including 1,460,756 genomes) for EHEC O157:H7 and Salmonella, respectively. Based on these two newly identified markers, combined with previously identified Cronobacter marker gene ygcB, multiplex PCR and multiplex TaqMan quantitative PCR methods were developed and established for the simultaneous detection of EHEC O157:H7, Salmonella, and Cronobacter. The specificity of the multiplex PCR and multiplex TaqMan qPCR was determined using 15 strains of Cronobacter, one strain of EHEC O157:H7, seven strains of Salmonella, and 100 strains of other common environmental bacteria and pathogens. The results showed that two methods demonstrated 100% specificity, both for multiple PCR and TaqMan qPCR. The detection limits of the multiplex PCR and TaqMan qPCR assays were 1 pg/μL and 0.5 pg/μL, respectively. Therefore, the newly developed methods are sensitive and reliable for the simultaneous identification of the three common food-borne pathogens.
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Data availability
All data supporting the findings of this study are available within the paper and its Supplementary Information. Furthermore, the original datasets (including in-house scripts) generated and/or analyzed during the current study are publicly available in the detect_rawdata repository, accessible via the GitHub platform: https://github.com/happywlu/detect_rawdata.
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Funding
This study is supported by the Natural Science Foundation of China (32201986), Natural Science Foundation of Hubei Province (2022CFB743), and Open Fund of Hubei Provincial Clinical Research Center for Precise Prevention and Treatment of Elderly Gastrointestinal Cancer (2024EGC-01).
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L. W. and L. Y. designed the research. H.N. Z., P. X., Z. L., and performed the research. X.L. C., R.H. Y., and L.L. Z. collected the data. Q.Y. Z., T.Y. L., and X.W. T. provided technical support and insights. P. X., Z. L., and L. W. analyzed the data. H.N. Z., P. X., and L. W. wrote the manuscript. L. W. revised the manuscript. All authors contributed to the article and approved the final manuscript.
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Zhang, H., Xiong, P., Lu, Z. et al. Novel marker genes for simultaneous detection of Salmonella, EHEC O157:H7, and Cronobacter. Sci Rep (2026). https://doi.org/10.1038/s41598-026-38990-x
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DOI: https://doi.org/10.1038/s41598-026-38990-x
