猪细小病毒VP2蛋白主要抗原域间接ELISA方法的建立  

The Establishment of Indirect ELISA Assay for the Major Antigenic Domain of Porcine parvovirus VP2 protein

作　　者：李斐[1] 李鹏[2] 郑其升[2] 于春梅 陈溥言[2] 

机构地区：[1]青岛农业大学生命科学学院,青岛266109 [2]南京农业大学农业部动物疫病诊断与免疫重点开放实验室,南京210095 [3]青岛宝依特生物制药公司,青岛266144

出　　处：《中国生物工程杂志》2007年第9期53-57,共5页China Biotechnology

基　　金：国家"863"计划资助项目(2001AA249012)

摘　　要：将已获得的含猪细小病毒VP2蛋白主要抗原域编码基因VP2 I重组酵母菌株在优化的条件下进行诱导表达,表达产物蛋白含量达227.6μg/ml,在此基础上以重组蛋白作为包被抗原初步建立了检测猪细小病毒抗体水平的间接ELISA方法,并对该方法进行了优化,结果表明抗原最佳包被浓度5.69μg/ml,而血清最佳稀释倍数为1∶80。阳性标准初步定为:OD待测样品>0.5,且OD待测样品/OD阴性血清>2.0。采用iVP2 I-ELISA对猪血清样品进行检测,结果显示iVP2 I-ELISA与HI试验的符合率为97.2%,与国外同类试剂盒的符合率达到91.2%。Followed the transformation of vp2 gene of PPV into the competent cell of Pichia pastoris, the vp2 protein of PPV was successfully expressed under the optimal expression conditions. The final concentration recombinant Vp2 protein was 227.6μg/ml,using the recombinant protein, an indirect ELISA assay for detection of PPV antibody was established . The optimal working circumstances for the antigen conditions（5. 69μg/ml） and the serum dilution were 1 ： 80, determined chess titration. The positive circumstances of this ELISA assay is OD490 value of the sample serum 〉 0.5, and the ratio of the tested serum OD490 to the negative serum is higher than 2.0. This ivp2-ELISA assay was compared with HA/HI and PPV ELISA kit of Cedi company, result shows the agreement ratio is 97.2% and 91.2% .

关 键 词：猪细小病毒 VP2蛋白 主要抗原域 酵母表达 间接ELISA 

分 类 号：Q819[生物学—生物工程]