鸡传染性腔上囊病病毒双夹心ELISA检测方法的建立  

Establishment of double sandwich ELISA for detecting infectious bursal disease virus

作　　者：马爱团[1] 钟秀会[1] 史万玉[2] 达来宝力格[2] 杨润德[2] 

机构地区：[1]河北农业大学中兽医学院,河北定州073000 [2]河北农业大学动物科技学院,河北保定071001

出　　处：《中国兽医科学》2008年第1期29-33,共5页Chinese Veterinary Science

基　　金：河北省教育厅博士基金项目(B2002211)

摘　　要：通过制备兔抗鸡传染性腔上囊病病毒(IBDV)、小鼠抗IBDV和绵羊抗兔免疫血清,并纯化绵羊抗兔辣根过氧化物酶标记抗体,采用方阵滴定法测定其最佳使用浓度,建立了IBDV双夹心ELISA检测方法。结果显示,包被抗体的最适稀释度为1∶160,兔抗IBDV的最佳稀释度为1∶200,酶标记抗体的最佳稀释度为1∶400。特异性和重复性试验结果表明,该方法特异性强,重复性好,与鸡新城疫、鸡传染性支气管炎和鸡减蛋综合征病毒抗原均不发生交叉反应。对人工感染病例的检测结果表明,建立的双夹心ELISA方法可用于临床快速诊断IBDV。Immune sera of rabbit/mouse anti-infectious bursal disease virus（IBDV） and sheep anti-rabbit were prepared,and immunoglobulins（IgGs） were purified from the three immune sera and labeled with HRP. Then,the optimal working concentrations of the IgGs were determined by using the chessboard titration crossing test and double sandwich ELISA based on the IgGs was established for the detection of IBDV. The results showed that the optimal working concentration of the IgGs was 1 ： 160,the optimal dilution of rabbit anti-IBDV IgG was 1 ： 200 and the optimal dilution of sheep anti-rabbit antibody labeled by HRP was 1 ： 400. Double sandwich ELISA had high specificity and reproducibility,which was confirmed by the repeating, blocking and crossing tests and it had no cross reaction with Newcastle disease virus（NDV）, infectious bronchitis virus（IBV） and eggs drop syndrome virus（EDSV）. It was confirmed by the diagnosis of the experimental cases that the established method could be used for rapid diagnosis of clinical IBDV.

关 键 词：鸡传染性腔上囊病病毒 HRP标记抗体 双夹心ELISA 

分 类 号：S852.659.4[农业科学—基础兽医学] R446.61[农业科学—兽医学]