Objective:To obtain monoclonal antibodies(McAb) against Staphylococcal enterotoxin C2(SEC2) and establish method for detecting SEC2.Methods:The secreted SEC2 from staphylococcus aureus was used as antigen to immune BALB/c mice. Monoclonal antibodies against SEC2 were prepared by normal hybridoma technique. By identifying the characters of McAbs, the quantitative detection ELISA test method were established and were preliminarily applied.Results:Four hybridmas producing antibodies against SEC2 were obtained. IgG isotypes of four McAbs were IgG1. Their binding site was different except two McAbs that shared the same binding site.The McAbs were proved to be specific for SEC2.The sandwich ELISA method had good specificity, sensitivity and reproducibility, and it was founded to be able to detect SEC2 at concentration from 0.5 to 20 ng/ml. Its recovery ranged between 97.8% and 101%,and CV value ranged between 2% and 5%.Conclusion:The prepared McAb against SEC2 can be used for SEC2 immunoassay. This work provides a basis for controlling the quality of JINPUSU and researching staphylococcal enterotoxin.