In order to develop detective kits of Seneca Valley virus(SVV),the 3ABC gene was expressed in E.coli BL21(DE3) and purified,and the purified protein was used to immunize rats.The polyclonal antibody against 3ABC protein was prepared,and its titer was determined by ELISA.Specific reaction was identified between the prepared polyclonal antibody and the expressed 3ABC protein by Western blot.The results showed that the 3ABC gene could be expressed in E.coli(mainly in inclusion body).Molecular weight of the recombinant 3ABC protein was about 55 ku.SDS-PAGE analysis showed that there was a clear single target band for the purified 3ABC protein.The titer of the polyclonal antibody was detected to be higher than 1∶64 000 by indirect ELISA.The polyclonal antibody could specifically recognize the recombinant 3ABC protein.In conclusion,SVV 3ABC gene could be successfully expressed in E.coli and the recombinant 3ABC protein had good immunogenicity.