Abstract:
Lumpy skin disease virus (LSDV) infection is a major socio-economic issue that seriously threatens the global cattle-farming industry. Here, a recombinant virus LSDV-ΔTK/EGFP, expressing enhanced green fluorescent protein (EGFP), was constructed with a homologous recombination system and applied to the high-throughput screening of antiviral drugs. LSDV-ΔTK/EGFP replicates in various kidney cell lines, consistent with wild-type LSDV. The cytopathic effect, viral particle morphology, and growth performance of LSDV-ΔTK/EGFP are consistent with those of wild-type LSDV. High-throughput screening allowed to identify several molecules that inhibit LSDV-ΔTK/EGFP replication. The strong inhibitory effect of theaflavin on LSDV was identified when 100 antiviral drugs were screened in vitro. An infection time analysis showed that theaflavin plays a role in the entry of LSDV into cells and in subsequent viral replication stages. The development of this recombinant virus will contribute to the development of LSDV-directed antiviral drugs and the study of viral replication and mechanisms of action.
摘要:
结节性皮肤病病毒(LSDV)感染是严重威胁全球养牛业的重大社会经济问题。这里,重组病毒LSDV-ΔTK/EGFP,表达增强型绿色荧光蛋白(EGFP),用同源重组系统构建并应用于抗病毒药物的高通量筛选。LSDV-ΔTK/EGFP在各种肾细胞系中复制,与野生型LSDV一致。细胞病变效应,病毒颗粒形态学,LSDV-ΔTK/EGFP的生长性能与野生型LSDV的生长性能一致。高通量筛选允许鉴定抑制LSDV-ΔTK/EGFP复制的几种分子。在体外筛选出100种抗病毒药物后,确定了茶黄素对LSDV的强抑制作用。感染时间分析表明,茶黄素在LSDV进入细胞和随后的病毒复制阶段中起作用。这种重组病毒的开发将有助于LSDV定向抗病毒药物的开发以及病毒复制和作用机制的研究。