猪伪狂犬病病毒变异株传代致弱JS18-150株的获得与鉴定  

Obtaining and identifying a mutated and passage-attenuated strain JS18-150 of porcine pseudorabies virus

作　　者：侯真真 张华伟 郝根喜 罗修鑫 宋文博 周明光 陈焕春 徐高原 HOU Zhenzhen;ZHANG Huawei;HAO Genxi;LUO Xiuxin;SONG Wenbo;ZHOU Mingguang;CHEN Huanchun;XU Gaoyuan(Wuhan Keqian Biology Co.,Ltd.,Wuhan 430206,China)

机构地区：[1]武汉科前生物股份有限公司,武汉430206

出　　处：《华中农业大学学报》2024年第5期178-185,共8页Journal of Huazhong Agricultural University

基　　金：湖北省重点研发计划项目(2023BBB177)。

摘　　要：为获得安全性良好、免疫原性高的变异株弱毒苗,采用鸡胚成纤维细胞(CEF)传代培养的方法对猪伪狂犬病病毒流行毒株JS18株进行传代,最终获得JS18-150株,随后通过PCR鉴定、测序、致病力试验和免疫效力试验对JS18-150株的体外生长特性、安全性和免疫原性进行初步研究。试验结果显示,在CEF上连续传代后,JS18株的gI、gE、US9基因缺失,获得的JS18-150株毒力已致弱,在CEF上适应性显著提高,与JS18株生长动力学相似;将JS18-150株以107.0TCID50/mL接种仔猪,所有仔猪均未观察到任何临床症状,且体温均未超过40.5℃;JS18-150株以106TCID50/mL接种仔猪后能保护仔猪有效抵御JS18株和HBZL05株的攻击。结果表明,JS18-150株对仔猪安全性良好,可作为PRV弱毒疫苗研制的候选毒株。Pseudorabies is an acute infectious disease of pigs,which is caused by the infection of pseu⁃dorabies virus(PRV).Pseudorabies is mainly prevented by vaccine,and PRV gene⁃deleted vaccine is the most widely used and effective.Since 2011,a new outbreak has occurred in many pig farms vaccinated with the Bartha⁃K61 vaccine and has spread to farms in most parts of the country,causing enormous economic losses to the development of our farms.The study found that the new outbreak was caused by PRV variants and showed that the commercially vaccine did not provide complete protection.Therefore,it is urgent to de⁃velop a new and efficient PRV vaccine.The prevalent strain JS18 of porcine pseudorabies virus was passaged with chicken embryo fibroblast(CEF)cells to obtain the attenuated vaccine with good safety and high im⁃munogenicity for mutated strain of porcine pseudorabies virus.JS18-150 strain was obtained.Preliminary studies were conducted on the in vitro growth characteristics,safety,and immunogenicity of JS18-150 strain through PCR identification,sequencing,pathogenicity testing,and immunogenicity testing.The re⁃sults showed that the gI,gE,and US9 genes of JS18 strain were deleted and the virulence of JS18-150 strain obtained was weakened after continuous passage on CEF.The adaptability on CEF of JS18-150 strain obtained was significantly improved.The growth kinetics of JS18-150 strain obtained was similar to that of the original JS18 strain.No clinical symptoms were observed in all piglets inoculated with 107.0 TCID50 JS18-150 strain obtained,and the rectal temperature did not exceed 40.5℃.Inoculation with 106.0 TCID50 JS18-150 strain obtained effectively protected piglets from the attacks of JS18 strain and HB⁃ZL05 strain.It is indicated that JS18-150 strain obtained has good safety for piglets and can be used as a candidate strain for the development of attenuated PRV vaccines.

关 键 词：猪伪狂犬病病毒 变异株 传代致弱 基因缺失 伪狂犬病疫苗 

分 类 号：S852.651[农业科学—基础兽医学]