Use of GFP-expressing influenza viruses for the detection of influenza virus A/H5N1 neutralizing antibodies
Use of GFP-expressing influenza viruses for the detection of influenza virus A/H5N1 neutralizing antibodies
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DOI：
10.1016/j.vaccine.2011.02.082
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发表时间：
2011-04-18
期刊：
影响因子：
5.5
通讯作者：
Osterhaus, Albert D. M. E.
中科院分区：
文献类型：
--
作者：
Rimmelzwaan, Guus F.;Verburgh, R. Joyce;Osterhaus, Albert D. M. E.
血凝抑制（HI）试验最常用于检测流感病毒抗体。然而，对于检测由感染或疫苗接种诱导的H5N1亚型禽流感病毒的抗体，HI测定不敏感。因此，病毒中和（VN）试验已成为检测针对这些病毒的人血清抗体的首选方法。然而，该测定需要第二次测定来检测残留病毒复制，这使得其进行起来费力并且不太适合于大量样品的高通量测试。在这里，我们描述了用于检测这些抗体的替代方法，其基于使用在感染靶细胞后表达绿色荧光蛋白（GFP）的报告病毒。产生携带多种抗原性不同的H5N1流感病毒的HA的表达GFP的病毒。事实证明，该方法易于实施，可以迅速进行。使用一组针对H5N1流感病毒产生的抗血清，将基于GFP表达病毒的测定与经典病毒中和测定和血凝抑制测定进行比较。一般而言，这些测定中获得的结果相关性良好。结论：基于报告病毒的检测是经典病毒中和检测的有吸引力的替代方法，适用于大型血清流行病学研究或临床试验中的疫苗有效性评估。(C)2011爱思唯尔有限公司保留所有权利。
The hemagglutination inhibition (HI) assay is used most commonly for the detection of antibodies to influenza viruses. However, for the detection of antibodies to avian influenza viruses of the H5N1 subtype either induced by infection or by vaccination, the HI assay is insensitive. Therefore, the virus neutralization (VN) assay has become the method of choice to detect human serum antibodies directed to these viruses. However, this assay requires a second assay for the detection of residual virus replication, which makes it laborious to perform and less suitable for high throughput testing of large numbers of samples. Here we describe an alternative method for the detection of these antibodies, which is based on the use of reporter viruses that express the green fluorescent protein (GFP) upon infection of target cells. GFP-expressing viruses were generated carrying the HA of a variety of antigenically distinct H5N1 influenza viruses. The method proved easy to perform and could be carried out rapidly. Using a panel of antisera raised against H5N1 influenza viruses, the assay based on GFP expressing viruses was compared with the classical virus neutralization assay and the hemagglutination inhibition assay. In general, the results obtained in these assays correlated well. It was concluded that the assay based on the reporter viruses is an attractive alternative for the classical virus neutralization assay and suitable for large sero-epidemiological studies or for the assessment of vaccine efficacy in clinical trials. (C) 2011 Elsevier Ltd. All rights reserved.