边缘无形体MSP2的原核表达及间接ELISA抗体检测方法的建立  

Prokaryotic Expression of Anaplasma marginale MSP2 Protein and Development of Indirect ELISA for Antibody Detection

作　　者：罗雨昕 赵敏 陈琬婷 邹予 咸承俊 王冬英[1,2,3,4] LUO Yuxin;ZHAO Min;CHEN Wanting;ZOU Yu;XIAN Chengjun;WANG Dongying(College of Animal Science and Technology,Guangxi University,Nanning Guangxi 530004,China;Guangxi Zhuang Autonomous Region Engineering Research Center of Veterinary Biologics,Nanning Guangxi 530004,China;Guangxi Key Laboratory of Animal Reproduction,Breeding and Disease Control,Nanning Guangxi 530004,China;Guangxi Colleges and Universities Key Laboratory of Prevention and Control for Animal Disease,Nanning Guangxi 530004,China)

机构地区：[1]广西大学动物科学技术学院,广西南宁530004 [2]广西壮族自治区兽用生物制品工程研究中心,广西南宁530004 [3]广西畜禽繁殖与疾病防控重点实验室,广西南宁530004 [4]广西高校动物疫病预防与控制重点实验室,广西南宁530004

出　　处：《现代畜牧科技》2024年第3期17-20,共4页Modern Animal Husbandry Science & Technology

基　　金：国家重点研发计划项目:广西大石山区优势特色产业提质增效技术集成与示范(2021YFD1100100)。

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。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∶2000,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.

关 键 词：边缘无形体 MSP2蛋白 原核表达 间接ELISA 

分 类 号：S852.6[农业科学—基础兽医学]