【摘要】 为建立可检测边缘无形体（Anaplasma marginale）的血清学检测方法，该研究根据GenBank收录的边缘无形体MSP2基因序列（登录号：EU526889），构建重组质粒pET28a-MSP2，原核表达获得pET28a-MSP2重组蛋白，将纯化后的蛋白作为抗原，建立间接ELISA检测方法。结果显示，该研究建立的间接ELISA方法，抗原最佳包被浓度、血清最佳稀释度和酶标二抗最佳工作浓度分别为8μg/mL、1∶400和1∶2 000，特异性、灵敏性和重复性良好。本试验成功表达并纯化了边缘无形体的MSP2蛋白，并建立了边缘无形体抗体间接ELISA方法，为边缘无形体的监测和诊断提供参考。更多还原

【Abstract】 In order to establish a serological assay for Anaplasma marginale,this study was conducted based on the gene sequence of A. marginale MSP2 collected by GenBank(Accession number:EU526889),the recombinant plasmid pET28a-MSP2 was constructed,and the pET28a-MSP2 protein was obtained and purified by prokaryotic expression.The purified protein was used as antigen to establish an indirect ELISA method.The results showed that the indirect ELISA method established in this study showed that the optimal concentration of antigen coated was 8μg/mL,the optimal serum dilution was 1∶400,and the optimal working concentration of the secondary antibody was 1∶2 000,which showed good specificity,sensitivity and repeatability.In this study,the MSP2 protein of A. marginale was successfully expressed and purified,and an indirect ELISA method for A. marginale antibody was established,which provided certain reference value for the monitoring,diagnosis for A. marginale.更多还原