摘要
将禽流感病毒（AIV）H5、H7、H9血凝素亚型标准血清中的IgG作为包被抗体，H5、H7、H9血凝素亚型单克隆抗体作为第二抗体．建立了双抗体夹心ELISA（AC—ELISA）方法．并优化了反应条件。结果表明：单抗和标准血清最适工作质量浓度分别为5、10mg／L，标准阳性血清于4℃包被12h．二抗作用条件为37℃作用1h，封闭条件为37℃作用1h，酶标抗体工作条件为1：5000稀释后，37C作用1h，底物作用条件为37℃作用10min。阴阳性抗原，临界值为0．274～0．332。本方法具有很高的特异性和敏感性。
Taking the IgG in standard serum of avian influenza virus H5, H7and H9 subtypes as coating antibody,while the monoclonal antibody of avian influenza virus H5, HTand H9 subtypes as the second antibody,the double antibody AC ELISA assay was performed and optimized to detect the nucleic acid of AIV. The result showed that the best condition as follows：the optimized mass concentration of monoclonal antibody and standard serum were 5 and 10 mg/L respectively;standard serum coating should be at the temperature of 4℃ for 12 h;the second antibody reaction should be at the temperature of 37℃ for 1 h;hermetization should be at the temperature of 37℃ for 1 h;enzyme labelled antibody should be diluted to 1 ： 5 000 ,and at the temperature of 37℃ for 1 h;the reaction of substrate should be at the temperature of 37℃ for 10 min; and the critical value to discriminate the negative antigen from the positive one is 0. 274-0. 332. The assay has a high specificity and sensitivity.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2009年第2期174-177,共4页
Chinese Journal of Veterinary Science
基金
科技部“十五”攻关资助项目(2004BA519A15)
