Establishment of A Detection Method for Listeria monocytogenes Based on Lateral Flow Immunity
Traditional microbial methods and real-time fluorescence polymerase chain reaction(PCR)methods could not fully meet the requirements of on-site testing in food safety assurance work for major events.Recombinant polymerase isothermal amplification(RPA)and lateral flow immunoassay(LFS)technology were combined together to detect Listeria monocytogenes in this study.The detection limit of the RPA-LFS in the pure bacteria,artificially contaminated meat products and artificially contaminated fresh fruits were 3.0×101 CFU/mL,3.0x101 CFU/g and 3.0×102 CFU/g respectively.A detection method for live bacteria was established by combining with propidium monoazide(PMA)treatment,which could inhibit the amplification of dead bacteria at a concentration level of 104 CFU/mL.The two methods and GB 4789.10-2016 were used for the detection of artificial contamination and actual samples,and it was found that PMA-RPA-LFS reduced false positives caused by dead bacteria.The established methods were applicable to food that has not been or has been heat treated.This study provided different choices for different types of food and new ideas for the detection of pathogenic bacteria in food safety assurance work during major events.
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