伪狂犬病病毒TK、gE双基因缺失传代致弱株的安全性及免疫效力研究  

Safety and immune efficacy of an attenuated pseudorabies virus variant by dual deletion of the TK and gE genes and serial passage

作　　者：朱来旭 许梦微[1,2,3] 张传健 郭容利[4] 王志胜 陈赛赛[2,3] 费荣梅[1] 王继春 ZHU Laixu;XU Mengwei;ZHANG Chuanjian;GUO Rongli;WANG Zhisheng;CHEN Saisai;FEI Rongmei;WANG Jichun(College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China;Institute of Veterinary Immunology&Engineering,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;National Research Center of Engineering and Technology for Veterinary Biologicals,Nanjing 210014,China;Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China)

机构地区：[1]南京农业大学动物医学院,江苏南京210095 [2]江苏省农业科学院动物免疫工程研究所,江苏南京210014 [3]国家兽用生物制品工程技术研究中心,江苏南京210014 [4]江苏省农业科学院兽医研究所,江苏南京210014

出　　处：《畜牧与兽医》2023年第2期50-55,共6页Animal Husbandry & Veterinary Medicine

基　　金：江苏省农业科技自主创新基金项目[CX(21)3130]。

摘　　要：为获得安全、有效的伪狂犬病病毒(pseudorabies virus,PRV)变异株弱毒疫苗,在前期研究的基础上,对PRV变异株TK、gE双基因缺失株(PRV LA1206株)进行传代致弱,在鸡胚成纤维细胞上连续传代75次,后经连续5代挑取单个病毒蚀斑克隆而得F_(80)代,毒种命名为PRV LA1206-80株,并对其进行安全性及免疫效力评价。生长动力学试验结果显示,PRV LA1206-80株与PRV LA1206株生长动力学相似。仔猪安全性试验结果显示,滴鼻接种1日龄PRV抗体阴性仔猪后所有仔猪均未出现发热及临床症状,表明该毒株对1日龄仔猪安全。攻毒保护试验结果显示,28~35日龄仔猪肌肉注射接种PRV LA1206-80株或PRV LA1206株7 d后进行变异株PRV AH02LA株滴鼻攻毒,可产生良好保护,所有仔猪均未出现体温反应、临床症状和排毒。而PRV Bartha K61株免疫猪7 d后攻毒,均出现了体温反应,其中1头发病,且鼻拭子样品均检出病毒。综上所述,相较于PRV LA1206株,在鸡胚成纤维细胞上连续传代后获得的PRV LA1206-80株已进一步致弱,对1日龄PRV抗体阴性仔猪安全,且该PRV TK、gE双基因缺失传代致弱株对PRV变异株的保护效果显著优于PRV Bartha K61株,是极具应用价值的PRV弱毒疫苗候选株。To obtain a safe and effective pseudorabies virus(PRV)variant attenuated vaccine,the PRV LA1206 strain was attenuated by the 75thserial passage on CEF cells.An F_(80)generation was obtained after five-round purification,and named the PRV LA1206-80 strain.The safety and immune efficacy of this PRV LA1206-80 strain were evaluated in this study.The results were as follow:The growth kinetics of the PRV LA1206-80 strain was similar to that of the PRV LA1206 strain.1-day-old PRV antibody-negative piglets were normal without fever or clinical symptoms of pseudorabies after intranasal inoculation of the PRV LA1206-80 strain,indicating that the strain was safe on 1-day-old piglets.The 28-35 days-old piglets were inoculated with the PRV LA1206-80 strain or the PRV LA1206 strain,and all were protected,and no viruses were shed after challenging with the PRV variant virulent strain AH02LA at day 7 post immunization.Meanwhile,when 28-35 days-old piglets were inoculated with the PRV Bartha K61 strain and subsequently challenged with PRV variant virulent strain AH02LA at day 7 post immunization,they all showed body temperature response and virus shedding by detection of their nasal swabs;one out of these piglets developed clinical symptoms of pseudorabies.In conclusion,the PRV LA1206-80 strain has been further attenuated,compared with that of the PRV LA1206 strain after serial passage on CEF cells,and it was safe on 1-day-old PRV antibody-negative piglets;at the same time,the PRV LA1206-80 strain showed a significantly better protection than the Bartha K61 strain.Therefore,the PRV LA1206-80 strain would be a very valuable PRV attenuated vaccine candidate strain.

关 键 词：伪狂犬病病毒 基因缺失 传代致弱 安全性 免疫效力 

分 类 号：S855.3[农业科学—临床兽医学]