流行性出血病病毒阻断ELISA抗体检测试剂盒的研制

Development on Blocking ELISA Kit for Antibodies against Epizootic Hemorrhagic Disease Virus

为研制流行性出血病病毒(epidemic hemorrhagic disease virus,EHDV)ELISA抗体检测试剂盒,用饱和硫酸铵浓缩EHDV灭活病毒作为包被抗原,以EHDV鼠源单克隆抗体作为阻断抗体,商品化的IgG-HRP羊抗鼠二抗作为检测抗体,通过优化检测试剂中各组分含量和反应程序,建立了一种EHDV阻断ELISA抗体检测方法。用研制的试剂盒与ID Screen参考试剂盒分别检测实验室保存的血清样本,结果发现该方法的特异性、敏感性与ID Screen参考试剂盒一致,二者符合率为97.00%。结果表明,研制的EHDV阻断ELISA抗体检测试剂盒可作为EHDV抗体检测的候选试剂盒,代替国外试剂盒使用,从而降低EHDV检测成本,这对该病防控具有重要意义。

In this study,a blocking ELISA kit for antibodies against epizootic hemorrhagic disease virus(EHDV)was established through optimization of contents and reaction procedures of each component by taking EHDV inactivated virus concentrated with saturated ammo nium sulfate as coating antigen,mouse monoclonal antibodies against EHDV as blocking antibodies and commercial IgG-HRP goat anti-mouse secondary antibodies as detection antibodies.Serum samples stored in the laboratory were tested by the established kit and ID Screen reference kit,respectively,it was found that the specificity and sensitivity of the two kits were consistent,with the coincidence rate of 97.00%.It was concluded that the established kit could be used as a candidate to replace any foreign kits,which could reduce the cost for detecting EHDV,and would play an important role in the prevention and control of the disease.