Chen Zhenhai, Yuan Fangfeng, Li Yanhua, Shang Pengcheng, Schroeder Robin, Lechtenberg Kelly, Henningson Jamie, Hause Benjamin, Bai Jianfa, Rowland Raymond R R, Clavijo Alfonso, Fang Ying
Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506, United States.
Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506, United States; Kansas State Veterinary Diagnostic Laboratory, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506, United States.
Virology. 2016 Oct;497:111-124. doi: 10.1016/j.virol.2016.07.003. Epub 2016 Jul 25.
A full-length cDNA infectious clone, pKS15-01-Clone, was constructed from an emerging Senecavirus A (SVA; strain KS15-01). To explore the potential use as a viral backbone for expressing marker genes, the enhanced green fluorescent protein (EGFP)-tagged reporter virus (vKS15-01-EGFP) was generated using reverse genetics. Compared to the parental virus, the pKS15-01-Clone derived virus (vKS15-01-Clone) replicated efficiently in vitro and in vivo, and induced similar levels of neutralizing antibody and cytokine responses in infected animals. In contrast, the vKS15-01-EGFP virus showed impaired growth ability and induced lower level of immune response in infected animals. Lesions on the dorsal snout and coronary bands were observed in all pigs infected by parental virus KS15-01, but not in pigs infected with vKS15-01-Clone or vKS15-01-EGFP viruses. These results demonstrated that the infectious clone and EGFP reporter virus could be used as important tools in further elucidating the SVA pathogenesis and development of control measures.
从一株新出现的猪水疱病病毒（SVA；KS15-01株）构建了全长cDNA感染性克隆pKS15-01-Clone。为了探索其作为表达标记基因的病毒骨架的潜在用途，利用反向遗传学技术构建了增强绿色荧光蛋白（EGFP）标记的报告病毒（vKS15-01-EGFP）。与亲本病毒相比，pKS15-01-Clone衍生病毒（vKS15-01-Clone）在体外和体内均能高效复制，并在感染动物中诱导产生相似水平的中和抗体和细胞因子反应。相反，vKS15-01-EGFP病毒生长能力受损，在感染动物中诱导产生的免疫反应水平较低。在所有感染亲本病毒KS15-01的猪的吻突背部和冠状带均观察到病变，但感染vKS15-01-Clone或vKS15-01-EGFP病毒的猪未出现病变。这些结果表明，感染性克隆和EGFP报告病毒可作为进一步阐明猪水疱病病毒发病机制和制定防控措施的重要工具。
