Analysis of Ebola virus and VLP release using an immunocapture assay
Analysis of Ebola virus and VLP release using an immunocapture assay
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DOI：
10.1016/j.jviromet.2005.02.015
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发表时间：
2005-07-01
影响因子：
3.1
通讯作者：
Aman, MJ
中科院分区：
文献类型：
--
作者：
Kallstrom, G;Warfield, KL;Aman, MJ
埃博拉病毒（EBOV）是一种新兴的病原体，是一种快速进行性出血热的病原体，具有高死亡率。目前还没有获得批准的埃博拉出血热疫苗或治疗方法。标准空斑试验是目前唯一可靠的病毒计数技术。有效的药物发现筛选以及靶标识别和验证需要简单且更快速的检测方法。本报告描述了一种快速ELISA的发展，测量病毒释放具有高灵敏度。该测定法直接从细胞培养上清液中检测埃博拉病毒和EBOV样颗粒（VLP），其中VP40基质蛋白用作抗原。使用该测定，确定EBOV核衣壳（NC）蛋白在VLP释放中的贡献。这些发现表明NC蛋白与包膜组分的组合对于VLP形成和释放是最佳的，这一发现对于用埃博拉VLP接种疫苗是重要的。此外，该检测方法可用于非生物防护环境中的替代模型，促进EBOV组装和出芽机制的基础研究以及药物发现研究。(c)2005 Elsevier B.V.保留所有权利。
Ebola virus (EBOV), an emerging pathogen, is the causative agent of a rapidly progressive hemorrhagic fever with high mortality rates. There are currently no approved vaccines or treatments available for Ebola hemorrhagic fever. Standard plaque assays are currently the only reliable techniques for enumerating the virus. Effective drug-discovery screening as well as target identification and validation require simple and more rapid detection methods. This report describes the development of a rapid ELISA that measures virus release with high sensitivity. This assay detects both Ebola virus and EBOV-like particles (VLPs) directly from cell-culture supernatants with the VP40 matrix protein serving as antigen. Using this assay, the contribution of the EBOV nucleocapsid (NC) proteins in VLP release was determined. These findings indicate that a combination of NC proteins together with the envelope components is optimal for VLP formation and release, a finding that is important for vaccination with Ebola VLPs. Furthermore, this assay can be used in surrogate models in non-biocontainment environment, facilitating both basic research on the mechanism of EBOV assembly and budding as well as drug-discovery research. (c) 2005 Elsevier B.V. All rights reserved.