A novel Ebola virus expressing luciferase allows for rapid and quantitative testing of antivirals.

A novel Ebola virus expressing luciferase allows for rapid and quantitative testing of antivirals.
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DOI:
10.1016/j.antiviral.2013.05.017
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发表时间:
2013-09
期刊:
影响因子:
7.6
通讯作者:
Feldmann, Heinz
Feldmann, Heinz
中科院分区:
医学2区
文献类型:
--
作者:
Hoenen, Thomas;Groseth, Allison;Callison, Julie;Takada, Ayato;Feldmann, Heinz

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埃博拉病毒(EBOV)引起严重的出血热,病死率高达90%,对此没有抗病毒治疗。抗病毒筛选受到以下事实的阻碍,即细胞病变效应的发展相对缓慢,而细胞病变效应是检测野生型EBOV感染的最简单手段。为了克服这个问题,我们产生了携带荧光素酶报告基因的重组EBOV。使用这种病毒,我们表明EBOV进入是快速的,在感染后2小时内可检测到病毒蛋白表达。此外,开发了基于发光的测定,以允许在48小时内进行高度灵敏的滴度测定。作为其在抗病毒筛选中的实用性的概念验证,我们使用该病毒来评估中和抗体和siRNA,其筛选时间比目前可用的野生型或重组病毒显著更快。这种重组病毒的可用性将允许针对EBOV的抗病毒药物的更快速和定量评估,以及EBOV生命周期细节的研究。
Ebola virus (EBOV) causes a severe hemorrhagic fever with case fatality rates of up to 90%, for which no antiviral therapies are available. Antiviral screening is hampered by the fact that development of cytopathic effect, the easiest means to detect infection with wild-type EBOV, is relatively slow. To overcome this problem we generated a recombinant EBOV carrying a luciferase reporter. Using this virus we show that EBOV entry is rapid, with viral protein expression detectable within 2 hours after infection. Further, luminescence-based assays were developed to allow highly sensitive titer determination within 48 hours. As a proof-of-concept for its utility in antiviral screening we used this virus to assess neutralizing antibodies and siRNAs, with significantly faster screening times than currently available wild-type or recombinant viruses. The availability of this recombinant virus will allow for more rapid and quantitative evaluation of antivirals against EBOV, as well as the study of details of the EBOV life cycle.
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