Visual Screening of Genetic Polymorphisms in Eae Gene of Escherichia coli O157:H7 with Single-Nucleotide Resolution by Arms Pcr-Mediated Lateral Flow Strip

Development of rapid, precise and fieldable detection methods for foodborne pathogen is one of the essential requirements in food safety and public health. In this research, the single nucleotide polymorphisms (SNPs) in the eae gene of Escherichia coli O157:H7 is well visually identified with the designed amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) mediated lateral flow strip (LFS). Allele specific primers were designed to discriminate the wild-type eae genes from the mutant-type with the single-nucleotide resolution in the simple visual LFS format. The single nucleotide variation in eae gene could be easily differentiated by the observation of optical signal on the T line of LFS without any devices. Assay performance results show that it has a high sensitivity and specificity with the single-nucleotide differentiation ratio as low as 0.1%. This genetic polymorphisms screening performance could enumerate complex genetic variation into a simple and direct yes/no readout, highlighting the ultra-easy SNP sensing mode and the simplicity of the results output for practical applications. This ARMS-PCR mediated LFS offers a straightforward, swift, and economical strategy for SNP identification with great potential using in evolution of bacterial resistance genes and viral evolution under different environmental stresses.