Agricultural Research Service Plum Island Animal Disease Center, Greenport, NY 11944, US.
Departments of Pathobiology and Veterinary Science, University of Connecticut, Storrs, CT 06269, USA.
Sci Rep. 2017 Apr 24;7:46747. doi: 10.1038/srep46747.
African swine fever is a contagious and often lethal disease for domestic pigs with a significant economic impact for the swine industry. The etiological agent, African swine fever virus (ASFV), is a highly structurally complex double stranded DNA virus. No effective vaccines or antiviral treatment are currently commercially available. We present here the development of a strain of ASFV that has been shown to retain its ability to cause disease in swine, efficiently replicate in swine macrophage and that is fluorescently tagged. The insertion of an EGFP cassette replacing the reading frames for two neighboring genes, MGF360-13L and MGF360-14L, in highly virulent field isolate Georgia/2007, did not affect virus replication in cell cultures and did not affect disease progression in swine, the natural host for ASFV. A virulent fluorescently tagged ASFV is a suitable tool to conduct pathogenesis studies in swine, study on virus-macrophage interaction and to run large scale screens that require a sensitive high throughput output. Utilizing an EGFP reporter system for observing ASFV replication and infectivity can circumvent the time and labor-intensive steps associated with viral antigen-based assays such as the observation of hemadsorption or cytopathic effect.
非洲猪瘟是一种传染性强、致死率高的疾病，对养猪业造成了重大的经济影响。其病原体为非洲猪瘟病毒（ASFV），是一种具有高度结构复杂性的双链 DNA 病毒。目前尚无有效的疫苗或抗病毒药物可供商业使用。在此，我们介绍了一株 ASFV 的开发情况，该病毒株能够在猪体内保持致病能力，在猪巨噬细胞中高效复制，并被荧光标记。在高致病性的田间分离株 Georgia/2007 中，插入一个 EGFP 盒，取代两个相邻基因 MGF360-13L 和 MGF360-14L 的阅读框，这不会影响病毒在细胞培养物中的复制，也不会影响 ASFV 的天然宿主猪的疾病进展。一株具有强毒力的荧光标记 ASFV 是一种适合在猪中进行发病机制研究、病毒-巨噬细胞相互作用研究以及进行大规模筛选的工具，这些筛选需要高灵敏度和高通量的输出。利用 EGFP 报告系统观察 ASFV 的复制和感染性，可以避免与病毒抗原检测相关的耗时费力的步骤，例如观察血吸附或细胞病变效应。
