应用原核表达的猪圆环病毒Ⅱ型Cap蛋白建立一种间接ELISA诊断方法  

Development of an indirect ELISA diagnostic method with the main antigentic region of PCV2 capsid protein expressed in E.coli

作　　者：林彦星[1] 孙彦伟 刘镇明[1] 陈靳松[3] 龚善中[1] 徐铮[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510640 [2]广东省动物防疫监督总所,广东广州510230 [3]华南农业大学动物科学学院,广东广州510640

出　　处：《中国预防兽医学报》2007年第3期217-221,共5页Chinese Journal of Preventive Veterinary Medicine

摘　　要：本试验在最佳诱导条件下获得猪圆环病毒Ⅱ型重组Cap蛋白的基础上,利用His Bind蛋白质纯化试剂盒对表达产物进行纯化,通过Western blot检测证明表达产物具有良好的抗原性。以纯化后的重组融合蛋白作为诊断抗原,对ELISA反应条件进行优化,初步建立了检测PCV2抗体的间接ELISA方法。用该法对广东、广西一些地区收集到的380份血清样品进行检测,检测结果阳性率为86.1%,从中随机取90份猪血清样品与国产商品化试剂盒检测结果对比,符合率为95.6%,表明本实验建立的间接ELISA方法具有较高的敏感性和特异性,适于大规模检测PCV2血清抗体的流行病学调查。An indirect enzyme-linked immunosorbent assay （ELISA） was developed based on a recombinant capsid protein of porcine circovirus type 2 （PCV2）. The recombinant capsid protein was expressed under the optimum induced condition, purified by a His Bind Purification Kit, and confirmed by Western blot assay. The indirect ELISA was used to test against 380 serum samples collected from Guangdong and Guangxi province and 86.1% samples were positive. Compared with commercial ELISA kits, the assay showed a correlation rate of 95.6 % based on the data obtained from 90 randomly picked serum samples. The results indicated that the indirect ELISA is not only sensitive and specific, but also suitable for large-scale epidemiological investigation for PCV2 infection.

关 键 词：PCV2 重组CAP蛋白 表达 纯化 ELISA 

分 类 号：S852.659.2[农业科学—基础兽医学] S854.44[农业科学—兽医学]