Abstract: Classical swine fever（CSF）caused by classical swine fever virus（CSFV），is a highly contagious and lethal viral disease in swine，for which vaccination remains a critical preventive strategy，antibody levels is an important indicator for evaluating the immune effect of immunized pig herds. In this study，the recombinant E2Y protein of CSFV was successfully expressed using a prokaryotic expression system. The purified E2Y protein served as a coating antigen，and an indirect ELISA method for detecting CSFV⁃specific antibodies was established through the systematic optimization of experimental conditions. Results indicated that the optimal coating concentration of the E2Y protein was 0.25 μg/mL. The assay demonstrated no cross⁃reactivity with positive sera against common swine viruses and exhibited high specificity，sensitivity，and reproducibility. The concordance between this method and a commercial ELISA kit reached 91.67% . This study successfully developed an indirect ELISA for CSFV antibody detection，providing a technical basis for further optimization and development of CSFV antibody detection reagents.

Key words: Classical swine fever virus, Antibody detection, Indirect ELISA method

摘要： 猪瘟（Classical swine fever，CSF）是由猪瘟病毒（Classical swine fever virus，CSFV）引 起的猪的一种高度传染性和致死性的病毒性疾病，疫苗接种是预防CSF的有效手段，抗体水平 是评价免疫猪群免疫效果的重要指标。该研究通过原核表达系统成功表达了CSFV重组蛋白 E2Y，将纯化的E2Y蛋白作为包被抗原，优化条件并建立了CSFV间接ELISA抗体检测方法。 试验结果表明，蛋白包被浓度为0.25 μg/mL，与常见猪病毒的阳性血清无交叉反应，具有良好 的特异性、灵敏性、可重复性，与商品化试剂盒一致性为91.67%。该研究成功建立了CSFV间 接ELISA抗体检测方法，为进一步优化和研制CSFV抗体检测试剂盒提供了技术参考。

关键词: 猪瘟病毒, 抗体检测, 间接ELISA方法