禽流感病毒M1抗体间接阻断ELISA检测方法的建立  

Establishment of an indirect-blocking ELISA for detection of antibody against matrix protein 1(M1)of avian influenza virus

作　　者：王兆鹏[1] 尚绪增[1] 刘思莹[1] 马波[1] 王君伟[1] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030

出　　处：《中国兽医科学》2009年第5期426-431,共6页Chinese Veterinary Science

基　　金：黑龙江省教育厅科学技术研究重大项目(10541Z004);黑龙江省科技攻关项目(GB04B504);黑龙江省青年基金项目(QC04C32)

摘　　要：以禽流感病毒(AIV)重组M1蛋白作为检测抗原,兔抗M1血清为阻断抗体,建立了检测AIVM1抗体的间接阻断ELISA方法。经筛选确定,抗原最佳包被浓度为0.78μg/mL,阻断抗体最佳稀释度为1∶15000,待检血清最佳稀释度为1∶10。用该ELISA方法对38份AIV阳性禽类样本和26份AIV阴性禽类样本进行检测,结果显示,该方法的敏感性为97.37%,特异性为96.15%。交叉试验证明该方法与新城疫等8种其他禽病阳性血清不发生交叉反应。用该ELISA方法对268份已知HI效价的临床样品进行检测,结果与HI的符合率达92%以上。表明,建立的间接阻断ELISA方法可用于AIV抗体的检测和禽流感的流行病学调查。An indirect blocking ELISA was established using recombinant avian influenza virus（AIV） matrix protein I（M1） as detection antigen, and using rabbit antiserum against the M1 protein as blocking antibody. The optimal concentration of the coating antigen was 0. 78 μg/mL, the optimal dilution of the blocking antibody was 1 ： 15 000,and the optimal dilution of the serum was 1 ： 10. 38 AIV positive samples and 26 AIV negative samples were detected by the indirect-blocking ELISA,indicating that the sensi- tivity of the indirect-blocking ELISA was 97. 37%, and the specificity was 96. 15%. Cross-reaction test showed that this method had no cross-reaction with positive sera against Newcastle disease virus,Marek＇s disease virus,infectious bronchitis virus, infectious bursal disease virus, infectious laryngotracheitis virus, duck plague virus, gosling plague virus, and goose paramyxovirus, respectively. The coincidence of the indirect-blocking ELISA with the hemagglutination inhibition（HI） test was over 92% for the detected 268 clinical samples. The results indicated that this method could be used for antibody detection and epidemiological investigation on AIV.

关 键 词：禽流感病毒 M1蛋白 间接阻断酶联免疫吸附试验 

分 类 号：S852.659.5[农业科学—基础兽医学] R446.61[农业科学—兽医学]