This article presents a new Polymerase Chain Reaction-Nucleic Acid Lateral Flow(PCR-NALF) method for the rapid detection of Listeria monocytogenes(LM) in food.Following isolation of DNA from the samples, PCR with two specific primers,one labeled with biotin and the other with digoxigenin,respectively,was performed. After applying the amplified product into a lateral flow strip that is coated with avidin and anti-digoxigenin antibodies, the test result can be interpreted according to the visible lines on the strip.The specifics were studied by testing a range of Listeria strains and other food relevant microorganisms.PCR products from other nonpathogenic Listeria,other microorganisms and the primer control(PCR without template DNA) were all negative,so this method can exactly differentiate LM from other common bacteria.And there is no observed difference(x~2 =0,P >0.05) when compared with classical microbiological detection method(GB/T 4789.30-2008).As a rapid,convenient,sensitive and specific assay method,the PCR-NALF can provide an effective means for LM detection in food samples.