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Development of a real‐time loop‐mediated isothermal amplification assay for the sensitive and rapid detection of Listeria monocytogenes

A real‐time loop‐mediated isothermal amplification (RealAmp) assay for the detection of Listeria was developed. The RealAmp assay, using primers specific for the hemolysin‐encoding hlyA gene, was verified using Listeria monocytogenes strains (n = 58) from different regions of the world. Both the sensitivity and specificity of the RealAmp assay were high. The RealAmp assay could detect 103 CFU ml−1 within 30 min. A comparative evaluation of the RealAmp assay, the API Listeria assay, and the real‐time PCR assay revealed that the RealAmp assay is simpler, faster, and has a higher specificity than the other two assays.