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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
【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。
【Abstract】 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.
【Key words】 porcine circovirus type 2; recombinant capsid protein; expression; purification; ELISA;
- 【文献出处】 中国预防兽医学报 ,Chinese Journal of Preventive Veterinary Medicine , 编辑部邮箱 ,2007年03期
- 【分类号】S854.43
- 【被引频次】29
- 【下载频次】300