Abstract: Two pairs of specific primers were composited based on the hlyA gene of L.monocytogenes and the 16sRNA gene of the genus Listeria.Optimal reaction system and condition of the SYBR GreenⅠfluorescent PCR were established, respectively, which could detect two specific genes.The reaction solution was verified by the method of gel electrophoresis.The result showed that two target fragments possessed different sizes and T_m valves which were amplfied corresponded two separate peaks.The standard curve based on the L.monocytogenes hlyA gene was established and the correlation coefficient was 0.996.The minimal detection limit was 89CFU/mL.Moreover, suspicious strain isolated from white clams son was tested by using this method so as to study the feasibility.The positive result was evaluated with the National Standard Methods (GB 4789.30-2010) .The verified result indicated that the positive result of SYBR GreenⅠfluorescent PCR matched with the GB 4789.30-2010 exactly.