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The high-performance of chemiluminescence immunoassays (CLIAs) in diagnosis has been gradually recognized in recent years, but their application in the diagnosis of classical swine fever (CSF) has not been reported. Here, a recombinant E2 (rE2) protein and a peroxidase-conjugated monoclonal antibody (MAb G5) were used to develop a competition-based chemiluminescence immunoassay (cCLIA) for rapid and accurate detection of E2-specific antibodies in pig serum. To evaluate the feasibility of cCLIA in the diagnosis of CSF, we developed a co