Polymerase Chain Reaction-Lateral Flow Strip for Detecting and Harboring .

and are common foodborne pathogens of global concern, particularly due to their antimicrobial resistance, notably to cephalosporins. This study aimed to evaluate a polymerase chain reaction-based lateral flow strip (PCR-LFS) assay for the detection of spp. and harboring the gene, which confers resistance to third-generation cephalosporins. Two duplex PCRs (dPCR) were established to detect -harboring (set 1) and -harboring (set 2). 600 bacterial isolates and raw pork mince spiked with -harboring and were used to evaluated. Both dPCR assays successfully detected -positive or strains, while strains lacking the gene showed no amplification. Non- and non- strains were PCR-negative unless they carried . The dPCR-LFS showed 100% validity including accuracy, sensitivity, specificity, positive predictive value, and negative predictive value for both or spp. harboring or lacking . The assay accurately detected target strains without cross-reactivity with other bacteria or antimicrobial resistance genes. Cohen's Kappa coefficient indicated perfect agreement ( = 1), reflecting the high reliability of the dPCR-LFS. The assay could detect as low as 25 CFU/mL for -positive and 40 CFU/mL for -positive in spiked raw pork mince. This assay is rapid, easy to interpret, and suitable for large-scale screening in surveillance programs.