埃博拉病毒病抗原反向间接血凝检测方法的建立

Development of a method for reverse indirect hemagglutination detection of Ebola virus disease antigen

为建立埃博拉病毒(Ebola virus,EBOV)抗原快速检测方法,本研究利用纯化后的埃博拉病毒样颗粒(EBOVVLPs)免疫马匹制备阳性血清,采用饱和硫酸铵沉淀法初纯血清中的IgG,后经过Protien G柱亲和层析法精纯IgG,利用精纯后的IgG致敏醛化的绵羊红细胞,建立反向间接血凝抗原检测方法,并对该方法进行最佳反应条件的优化。将该方法用于不同批次制得的已知病毒滴度埃博拉假病毒血凝效价检测,检测结果显示:病毒滴度的高低与血凝效价检测结果呈现良好的相关性。本研究成功建立埃博拉病毒抗原反向间接血凝检测方法,该方法简单、方便、快捷,无需特殊的仪器设备,1h内便可报告检测结果,在普通实验室中便可进行操作。此外,该方法也适用于埃博拉病毒感染及疫病暴发后临床样品的定性分析。

In order to establish a rapid detection method for Ebola virus(EBV)antigen,in this study,purified Ebola virus-like particles(EBOV-VLPs)were used to immunize horses for preparing positive serum.The saturated ammonium sulfate precipitation method were employed for purification. The IgG was purified by Protien G column affinity chromatography,and the purified IgG was used to sensitize the hydroformylated sheep erythrocytes,and the reverse indirect hemagglutination antigen detection method was established.The reaction condition of the method was optimized. The method was applied to the detection of the Ebola pseudovirus with known viral hemagglutination titer in different batches.It indicates that the results were highly consistent and showed a close relativity.This study successfully established a reverse indirect hemagglutination test method for Ebola virus antigen.The method is simple,convenient and fast,and requires no special equipment.The test results can be reported within 1hour and can be operated in an ordinary laboratory. In addition,the method is also applicable to the qualitative analysis of clinical samples after EBOV infection and disease outbreak.