基于评估大熊猫源CDV/CPV病毒样颗粒疫苗抗体的ELISA方法建立及应用

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中图分类号：S852.655 文献标志码：A 文章编号：1004-390X（2025）03-0068-10

Abstract： [Purpose] To establish two indirect ELISA methods for detecting the specific antibody levels， after immunization with giant panda-derived canine distemper virus （CDV）/canine parvovirus （CPV） virus-like particle （VLP） vaccines. [Methods] Giant panda-derived CDV was used as a base material， plasmids containing the major antigenic epitopes of F protein and H protein were constructed for prokaryotic expression. The recombinant proteins were expressed in Escherichia coli BL21 （DE3） strain; after purification， dialysis and refolding， the reactogenicity of the recombinant proteins were detected.FH fusion protein and giant panda-derived CPV VP2 protein were used as ELISA coating antigens， the antigen mass concentration， serum dilution， and other assay conditions were optimized，and then， the specificity，sensitivity，and repeatability were assessed.The two ELISA methods， as well as a commercial CDV/CPV antibody detection kit， were used to measure the specific antibody levels in the serum of CDV/CPV VLP-immunized dogs and evaluate the agreement between the methods.[Results]A fusion protein was successfully obtained. For FH fusion protein as the coating antigen， the optimal coating mass concentration was 8μg/mL ，with the best serum dilution at 1：800 ； for VP2 protein as the coating antigen， the optimal coating mass concentration was 4μg/mL ， with the best serum dilution at 1：400 . Two indirect ELISA methods for detecting giant panda-derived CDV and CPV specific antibodies were successfully established， showing good specificity， sensitivity，and repeatability， with sensitivity superior to that of the commercial antibody detection kit. [Conclusion] The two indirect ELISA methods established in this study can accurately reflect the immunization efect of the giant panda-derived CDV/CPV VLP particle vaccines， and they possess significant clinical application value.

Keywords： giant panda; canine distemper virus; canine parvovirus; virus-like particle vaccine; ELISA

大熊猫（Ailuropodamelanoleuca）是全球珍稀且易危物种，其种群繁育和保护具有重要的生态学和生物学意义。（剩余13909字）