Influenza virus: a novel method to assess viral and neutralizing antibody titers in vitro
Influenza virus: a novel method to assess viral and neutralizing antibody titers in vitro
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DOI：
10.1016/s0022-1759(99)00034-4
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发表时间：
1999-05-27
影响因子：
2.2
通讯作者：
Kopf, M
中科院分区：
文献类型：
--
作者：
Bachmann, MF;Ecabert, B;Kopf, M
本报告描述了一种新的体外测定流感病毒滴度和病毒中和抗体水平的方法。为了测定病毒滴度，将流感病毒的一系列稀释物与mdck细胞一起孵育，并在甲基纤维素覆盖层下在24孔板中培养48小时。将细胞固定、通透化，并用血凝素（HA）特异性单克隆抗体和过氧化物酶标记的第二阶段抗体进行染色。该试验的灵敏度比使用新鲜鸡血的常规血凝试验高100-1000倍。为了测定流感病毒的中和活性，将病毒样品与一系列稀释的抗体孵育，并按照上述相同的程序在96孔板中评估剩余病毒活性。该试验可以区分IgM和IgG抗体滴度，并且比使用新鲜鸡血的经典血凝抑制试验灵敏约5-10倍。(C) 1999 Elsevier Science B.V.版权所有
The present report describes novel in vitro assays to determine influenza virus titers and virus neutralizing antibody levels. For determination of viral titers, serial dilutions of influenza virus were incubated with MDCK-cells and cultured for 48 h under a methylcellulose overlay in 24 well plates. Cells were fixed, permeabilized and stained with a monoclonal antibody specific for hemagglutitin (HA) and a peroxidase labelled second stage antibody. The sensitivity of the assay was 100-1000 times greater than a conventional hemagglutination test using fresh chicken blood. For determination of influenza virus neutralizing activity, viral samples were incubated with serial dilutions of antibody and residual viral activity was assessed in 96 well plates by the same procedure as described above. This assay made it possible to distinguish between IgM and IgG antibody titers and was about 5-10 fold more sensitive than a classical hemagglutination inhibition assay using fresh chicken blood. (C) 1999 Elsevier Science B.V. All rights reserved.