Host-encoded reporters for the detection and purification of multiple enveloped viruses.
Host-encoded reporters for the detection and purification of multiple enveloped viruses.
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DOI：
10.1016/j.jviromet.2010.04.002
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发表时间：
2010-08
影响因子：
3.1
通讯作者：
Seed B
中科院分区：
文献类型：
--
作者：
Ketteler R;Tomov V;Neunkirchner A;Xie Q;Pickl WF;Seed B
病毒复制的宿主细胞因子的识别为新的抗病毒疗法的开发带来了巨大的希望。最近，高通量筛选方法已经成为识别治疗干预的候选宿主因素的有力工具。适合大规模自动化筛查的检测系统的开发对于具有高致病潜能的新病毒尤其重要，因为在短时间内可以开发有限的生物学信息。本报告提出了一种通用的酶报告系统，用于检测和鉴定多囊膜病毒，而不依赖于病毒的工程。取而代之的是，报告酶通过靶向产生细胞中的脂质微域而被结合到病毒颗粒中。该方法允许通过灵敏、廉价和自动化的酶分析来检测各种人类致病包膜病毒。基于磁珠的捕获方法可以快速高效地提纯标记的病毒。该方法允许在不事先参考其序列的情况下对包膜病毒进行一般检测。
The identification of host cell factors for virus replication holds great promise for the development of new antiviral therapies. Recently, high-throughput screening methods have emerged as powerful tools to identify candidate host factors for therapeutic intervention. The development of assay systems suitable for large-scale automated screening is of particular importance for novel viruses with high pathogenic potential for which limited biological information can be developed in a short period of time. This report presents a general enzymatic reporter system for the detection and characterization of multiple enveloped viruses that does not rely on engineering of the virus. Instead, reporter enzymes are incorporated into virus particles by targeting to lipid microdomains in producer cells. The approach allows a variety of human pathogenic enveloped viruses to be detected by sensitive, inexpensive and automatable enzymatic assays. Tagged viruses can be purified quickly and efficiently by a magnetic bead-based capture method. The method allows general detection of enveloped viruses without prior reference to their sequence.
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