H5亚型高致病性禽流感病毒抗原捕捉ELISA诊断方法的建立  

Development of antigen capture ELISA for detecting H5 subtype high pathogenic avian influenza virus

作　　者：曲站红[1] 邓国华[1] 王桂芹[1] 张立春[1] 田国彬[1] 于康震[2] 陈化兰[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所农业部动物流感重点开放实验室/兽医生物技术国家重点实验室 [2]中国兽医药品监察所,北京100081

出　　处：《中国预防兽医学报》2008年第1期68-71,80,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家十五科技攻关项目资助(2004BA519A54)

摘　　要：本研究在已有H5亚型禽流感病毒特异性单抗基础上,建立了以多抗作为包被抗体、单抗作为检测抗体的抗原捕捉ELISA方法。通过对各个反应条件进行优化,获得的最佳工作条件为:羊血清1∶1600倍稀释;单抗1∶10000稀释;酶标抗体最适工作浓度为1∶5000倍稀释;单抗反应时间1.5h;酶标抗体作用时间1.5h。特异性试验和敏感性试验结果表明:该方法具有良好的特异性和敏感性,可用于H5亚型高致病性禽流感病毒的检测。An antigen capture ELISA was developed for detection of H5 HPAIV using polyclonal antibody as capture antibody and H5 specific monoclonal antibody as detecting antibody. The assay was optimized to use following antibody dilutions： capture antibody at 1：1600, H5 subtype specificity McAb at 1：10000 and the enzyme-labeled antibody at 1：5000. The optimal incubation time for the detecting antibody and conjugate were one and half an hour, respectively. This method had high specificity and sensi- tivity and could be used for diagnosing H5 subtype HPAIV.

关 键 词：禽流感病毒 H5亚型 ELISA 

分 类 号：S852.659[农业科学—基础兽医学] S854.43[农业科学—兽医学]