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. 2021 Jan 21;11:1881. doi: 10.1038/s41598-021-81312-6

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Primer screening and specificity test (a) Primer selection: Lane 1, 10 and 19: 100 bp molecular marker, Lane 2 and 3: rfbE F1/R1; lane 4 and 5: rfbE F2/R2; lane 6 and 7: rfbE F3/R3; lane 8 and 9: rfbE F4/R4; lane 11 and 12: fliC F1/R1; lane 13 and 14: fliC F2/R2; lane 15 and 16: fliC F3/R3; lane 17 and 18: fliC F4/R4; lane 20 and 21: stx1 F1/R1; lane 22 and 23: stx1 F2/R2; lane 24 and 25: stx2 F1/R1; lane 26 and 27: stx2 F2/R2. Specificity Assay (b) Amplification based on the rfbE F1/R1 primer set, (c) Amplification based on the fliC F2/R2 primer set and (d) Amplification based on the stx2 F1/R1 primer set—with RPA-AGE using E. coli O157:H7, Staphylococcus aureus, Shigella flexneri, Salmonella thyphimurium, E. coli K-12, Enterococcus faecalis, non-template control (NTC). M molecular marker, T template, N non-template control.