猪圆环病毒2型抗体检测间接ELISA方法的建立  

Development of indirect ELISA using recombinant PCV2 ORF2 protein as antigen of porcine circovirus 2

作　　者：丛丽媛[1] 蒋智勇[2] 张春红[2] 陈德坤[1] 宋长绪[2] 

机构地区：[1]西北农林科技大学,陕西杨凌712100 [2]广东省农业科学院兽医研究所,广东广州510640

出　　处：《中国兽医学报》2008年第8期892-896,共5页Chinese Journal of Veterinary Science

基　　金：国家食品安全重大专项(2001BA804A31);广东省农业科技攻关(2003B21404,20052490100,2006B20301013);广东省自然科学基金资助项目(04002083);广东省自然科学基金重点项目(06105311)

摘　　要：用纯化的重组猪圆环病毒2型(PCV2)ORF2蛋白作为包被抗原,对各种条件进行优化(如抗原的包被、血清及酶标二抗作用时间、底物的选择等),建立了检测猪圆环病毒2型(PCV2)抗体的间接酶联免疫吸附试验(ELISA)的方法。结果显示,重组抗原最适包被浓度为0.5mg/L,最适包被条件为4℃过夜,血清稀释度1∶160,酶标二抗工作浓度1∶7500,待检测血清和酶标二抗的反应时间均为37℃,45min,底物37℃显色10min。阻断试验表明,建立的间接ELSA对PCV2抗体具有很好的特异性。板内和板间重复试验显示,同1份血清板内各孔的变异系数均值为2.78%,板间同一份血清D值变化不大,表明重复性好。The purified PCV2 ORF2 protein was as the antigen to coat a 96-well plate,and the following factors affected the experiments were optimized.The results demonstrated that optimal concentration of recombinant ORF2 protein for coating was 0.5 mg/L,the optimal coating condition of recombinant ORF2 protein for ELISA was incubated at 4℃ overnight,the dilution of serum sample was 1∶160,the work concentration of HRP-labeled rabbit anti-porcine IgG was 1∶7 500,the best incubation period and temperature for both the ...

关 键 词：猪圆环病毒2型 间接ELISA 重组蛋白 

分 类 号：S854.43[农业科学—临床兽医学]