Objective To get expression of Bordetella bronchiseptic recombinant protein DNT1 and establish an indirect ELISA for detection of recombinant protein DNT1. Method According to a pair of specific primers designed to amplify DNT gene of Bordetella bronchiseptica in swines promulgated by Genbank，the fragment of the target gene was amplified by PCR and then constructed with prokaryotic expression vector PET-28a ( + ) . The recombinant expression plasmids were transfected into BL21(DE3) strain. The optimal concentration of coated antigen recombinant protein DNT1 and serum dilution was determined to develop the ELISA technique. Results DNT1 gene of Bordetella bronchiseptica was successfully cloned and expressed. The SDS-PAGE and Western blot analysis unfolded the excellent immunogenicity of the recombinant proteins which were used as coating antigens to develop the ELISA method for Bb-specific antibody diagnosis.The peridium consistency of the recombinant protein DNT1 determined in this experiment was 6. 25 μg /mL，and the optimal testing serum dilution was 1:100. Conclusion The development of DNT1 indirect ELISA offers a simple and practical way for monitoring antibody of Bb，and provides much information for laying a basis for development of a Bb diagnosis kit.