Development of a homogeneous screening assay for automated detection of antiviral agents active against severe acute respiratory syndrome-associated coronavirus
Development of a homogeneous screening assay for automated detection of antiviral agents active against severe acute respiratory syndrome-associated coronavirus
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DOI：
10.1016/j.jviromet.2005.05.010
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发表时间：
2005-10-01
影响因子：
3.1
通讯作者：
Hertogs, K
中科院分区：
文献类型：
--
作者：
Ivens, T;Van den Eynde, C;Hertogs, K
2003年爆发的严重急性呼吸综合征相关冠状病毒（SARS-CoV）的严重程度和全球传播突出了新出现的病毒性疾病对人类健康构成的风险，并强调需要使用特定的治疗剂，而不是依赖现有的广泛有效的抗病毒化合物。快速筛选试验的发展对抗病毒药物的发现至关重要。因此，建立了抗sars - cov药物筛选系统，在初始筛选阶段评估化合物的效力、特异性和细胞毒性。对细胞系进行改造，使其组成性地表达增强型绿色荧光蛋白（EGFP），并用于检测(1)SARS-CoV感染试验中的抗病毒效力；(2)猪冠状病毒传染性胃肠炎病毒（TGEV）抗病毒特异性试验；(3)在没有病毒攻击的情况下进行相同的细胞毒性试验。该检测系统在检测设置后操作最少，便于自动读出，并最大限度地降低与危险病毒相关的风险。通过对3388个小分子化合物的筛选，证明了该系统在药物发现中的适用性。结果表明，这些方法可用于高通量筛选鉴定选择性抗SARS-CoV活性抑制剂。(c) 2005 Elsevier B.V.版权所有
The severity and global spread of the 2003 outbreak of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) highlighted the risks to human health posed by emerging viral diseases and emphasized the need for specific therapeutic agents instead of relying on existing broadly active antiviral compounds. The development of rapid screening assays is essential for antiviral drug discovery. Thus, a screening system for anti-SARS-CoV agents was developed, which evaluated compound potency, specificity and cytotoxicity at the initial screening phase. Cell lines were engineered to constitutively express an enhanced green fluorescent protein (EGFP) and used to detect (1) antiviral potency in SARS-CoV infection tests; (2) antiviral specificity in tests using the porcine coronavirus transmissible gastroenteritis virus (TGEV); and (3) cytotoxicity in the same assays without virus challenge. The assay system involves minimal manipulation after assay set-up, facilitates automated read-out and minimizes risks associated with hazardous viruses. The suitability of this assay system in drug discovery was demonstrated by screening of 3388 small molecule compounds. The results show that these assays can be applied to high-throughput screening for identification of inhibitors selectively active against SARS-CoV. (c) 2005 Elsevier B.V. All rights reserved.