Prokaryotic expression of the structural protein S of porcine epidemic diarrhea virus and establishment of an indirect ELISA for an detection of antibody against porcine epidemic diarrhea

Abstract:

[Objective] To develop a rapid, specific and sensitive method for detecting the serum antibodies against porcine epidemic diarrhea virus (PEDV). [Methods] The major S protein antigenic region were expressed after analyzing the site antigenic of PEDV S protein by bioinformatics softwares and the recombinant protein was detected by SDS-PAGE and Western-blot. Furthermore, indirect ELISA was developed by using the recombinant protein as coating antigen and though optimizing conditions, specificity and reproducibility tests. [Results] The recombinant protein was expressed successfully, the recombinant S protein could react with PEDV positive serum specifically, and the indirect ELISA method for serum antibodies against PEDV was developed successfully. Both the intro-batch and inter-batch variation coefficient were lower than 10%. The developed indirect ELISA method compared with commercialization of PEDV antibody detection kit and Western-blot, the coincidence rate was 86.67% and 88.89% respectively. [Conclusion] The established indirect ELISA could be used for detecting PEDV antibodies.