Abstract: Listeria monocytogenes is a foodborne pathogenic bacteria which can cause human and animal diseases.Monitoring and testing Listeria monocytogenes in food is very important. In order to determine the minimum detection limit of real-time fluorescent PCR in the detection of food borne Listeria monocytogenes and the detection efficiency in different food types, the limit of detection of real-time fluorescent PCR method for Listeria monocytogenes were verified in this paper. Listeria monocytogenes was added in four kinds of foods, namely soybean meal, biscuit, lettuce and pork, and was detected by real-time fluorescent PCR method at differrent time points.Through comparing and analyzing with the national standard method, the real-time fluorescent PCR method has the advantages of fast and high sensitivity, and can be applied in rapid detection. However, considering that the dead bacteria can cause false positive problems, the national standard method can seperate the contaminated bacterial strain for further research, it is recommended to recheck and confirm the positive samples using national standard method after PCR detection.