Center for Computational and Integrative Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA.
J Virol Methods. 2010 Aug;167(2):178-85. doi: 10.1016/j.jviromet.2010.04.002. Epub 2010 Apr 23.
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.
宿主细胞因子的鉴定为开发新的抗病毒疗法带来了巨大的希望。最近,高通量筛选方法已成为鉴定治疗干预候选宿主因子的有力工具。适合大规模自动化筛选的测定系统的开发对于具有高致病性潜力的新型病毒尤为重要,因为对于这些病毒,在短时间内很难获得有限的生物学信息。本报告介绍了一种通用的酶报告系统,用于检测和表征多种包膜病毒,该系统不依赖于病毒的工程改造。相反,报告酶通过靶向生产细胞中的脂质微区而被掺入病毒颗粒中。该方法可以通过灵敏、廉价和自动化的酶测定法检测多种人类致病性包膜病毒。标记的病毒可以通过基于磁珠的捕获方法快速有效地纯化。该方法允许在不事先了解其序列的情况下进行通用的包膜病毒检测。