中国动物传染病学报2025，Vol.33Issue(2)：26-34,9.
柯萨奇病毒A组16型感染性克隆的构建及免疫原性评价
Construction of CVA16 Infectious Clones and Evaluation of Immunogenicity
摘要
Abstract
Coxsackievirus A group 16(CVA16)is one of the major pathogens causing epidemics and outbreaks of hand,foot and mouth disease,which poses severe challenges to public health.The cDNA infectious clone of CVA16 was rescued in vitro,and its infectivity,serial passage stability and in vivo immunogenicity were studied to provide a basis for the development of CVA16 vaccine.In this study,the eGFP reporter gene system was used to build the CVA16 minigenome platform,which laid the foundation for the construction of CVA16 infectious clones.The infectious clone of CVA16 was further rescued by the reverse genetic operating system and examined for its infection ability,plaque morphology and serial passage stability.Finally,the immunogenicity of CVA16 infectious clone was evaluated in mice.The constructed full-length cDNA plasmid of CVA16 was confirmed by restriction endonuclease double digestion to be a target band of about 7500 bp and a vector band of about 3500 bp.The constructed clone was verified to be completely consistent with the expected sequence by whole gene sequencing.Observation on cell morphology showed typical enterovirus-induced cytopathic changes in CVA16 group.After continuous passages in cell cultures,CVA16 still showed constant infectivity and plaques looked normal.Afterwards,mice were inoculated with the CVA16 infectious clone.As a result,higher titers of specific antibodies and neutralizing antibodies were produced and the titer of neutralizing antibodies induced by ultra-concentrated CVA16 was twice higher that of the stock solution immunized group.Taken together,the CVA16 infectious clone obtained by reverse genetic manipulation technology stably proliferated in vitro and stimulated high titer of neutralizing antibodies in mice.This study laid the foundation for in-depth research on the gene function,infection,pathogenic mechanism and vaccine development of the virus.关键词
柯萨奇病毒A组16型/反向遗传/感染性克隆/中和抗体Key words
Coxsackievirus A group 16/reverse genetics/infectious clone/neutralizing antibody分类
医药卫生引用本文复制引用
李巨银,葛君,马超,张建辉,闫伟,孙娇娇,李建强..柯萨奇病毒A组16型感染性克隆的构建及免疫原性评价[J].中国动物传染病学报,2025,33(2):26-34,9.基金项目
泰州市科技支撑计划(农业)项目(TN202002,NSFZP202204) （农业）
江苏高校"青蓝工程"项目(苏教师函[2022]29号) （苏教师函[2022]29号）
