鸽副黏病毒灭活疫苗候选株aYQ的生物学特性研究  

Biological Characteristics of Inactivated Vaccine Candidate aYQ of Pigeon Paramyxovirus

作　　者：赵文奇 杨志远[2] 林健[2] 陈光华[3] 杨龙峰 赵际成 段会娟[2] 丁国杰 马会籽 王小蕾[2] 刘立新[2] 田方杰 潘洁[2] 程水生 孙晔 蒋桃珍[3] 刘月焕[2] ZHAO Wen-qi;YANG Zhi-yuan;LIN Jian;CHEN Guang-hua;YANG Long-feng;ZHAO Ji-cheng;DUAN Hui-juan;DING Guo-jie;MA Hui-zi;WANG Xiao-lei;LIU Li-xin;TIAN Fang -jie;PAN Jie;CHENG Shui-sheng;SUN Ye;JIANG Tao-zhen;LIU Yue-huan(College of Animal Science and Technology,Beijing University of Agriculture,Beijing,102206,China;Institute of Animal Husbandry and Veterinary Medicine,Beijing Academy of Agricultural and Forestry Sciences,Beijing,100097,China;China Institute of Veterinary Drug Control,Beijing,100081,China;Animal Diseasees Prevention and Control Center of Yanqing District,Beijing,102100,China;Beijing Zhonghai Biotech Co., Ltd, Beijing, 100081,China)

机构地区：[1]北京农学院动物科学技术学院,北京102206 [2]北京市农林科学院畜牧兽医研究所,北京100097 [3]中国兽医药品监察所,北京100081 [4]北京市延庆县动物疫病预防控制中心,北京102100 [5]北京中海生物科技有限公司,北京100081

出　　处：《动物医学进展》2019年第1期36-41,共6页Progress In Veterinary Medicine

基　　金：北京市农林科学院科技创新能力建设专项项目(KJCX20161503);北京市农林科学院畜牧兽医研究所项目(XMS201703)

摘　　要：评价鸽副黏病毒(PPMV)致弱毒株aYQ的生物学特性,为研制鸽副黏病毒灭活疫苗奠定基础。将PPMV aYQ株经SPF鸡胚连续传代,建立种子批。测定各代次毒种的血凝效价(HA)和鸡胚半数感染量(EID50),并进行纯净性检验;测定毒种鸡胚最小致死量的平均死亡时间(MDT)、1日龄鸡脑内接种致病指数(ICPI)和6周龄鸡静脉接种致病指数(IVPI);测定毒种对鸡胚和鸽的毒力、遗传稳定性和抗原相关性。PPMV aYQ株经SPF鸡胚传代至F10,HA效价为1∶256～1∶512,病毒含量为10-8.5 EID50/0.1mL～10-8.9 EID50/0.1mL,毒种各代次均无细菌、霉菌、支原体以及外源病毒污染;PPMV aYQ株的ICPI和IVPI结果均为0,MDT测定结果大于120h;以103倍稀释的PPMV aYQ株病毒液接种10日龄SPF鸡胚96h无死亡,无眼观病变。以106EID50/羽的剂量人工感染鸽,未见异常症状;PPMV aYQ株各代次F基因均未发生基因突变,F蛋白裂解位点氨基酸序列均为-112GRQGRL 117-;交叉血凝抑制试验和交叉中和试验结果表明,PPMV aYQ株与NDV La Sota株间存在明显的抗原性差异。鸽副黏病毒弱毒株aYQ能够在SPF鸡胚上稳定传代,且生物学特性稳定,免疫原性良好,可以作为鸽源副黏病毒灭活疫苗的毒种。The objective of this study was to evaluate the biological characteristics of the aYQ strain of pigeon paramyxovirus and to lay the foundation for the development of the pigeon-paramyxovirus inactivated vaccine.PPMV aYQ strain were passaged continuously in SPF chicken embryos to establish a seed batch.HA and EID 50 were determined and the purity of each strain was tested;The virulence of the aYQ strain was evaluated by MDT,ICPI and IVPI.Pathogenicity of aYQ strain to chicken embryos and pigeons,genetic stability and antigen relevance were determined.PPMV aYQ strain was passaged to 10 generations in SPF chicken embryos in the absence of bacteria, mold, mycoplasma and exogenous viruses.The HA titers of various generation viruses were 1∶256-1∶512 and the virus contents were 10^-8.5 -10^-8.9 EID 50 /0.1 mL.Both ICPI and IVPI of PPMV aYQ strain were 0,and the MDT was more than 120 h; The 10-day-old SPF chicken embryos inoculated with 10^3-fold diluted PPMV aYQ strain survived at 96 h post inoculation.Pigeons which were infected artificially at a dose of 10 6EID 50 with PPMV aYQ strain showed no apparent clinical symptoms;The results of sequence analysis showed that there was no gene mutation in F gene of PPMV aYQ strain,and the deduced amino acid sequences were -112 GRQGRL 117-.The results of cross-hemagglutination inhibition test and cross-neutralization test showed that there was obvious antigenic difference between PPMV aYQ strain and NDV La Sota strain.The aYQ strain can be passaged in SPF chicken embryos with stable genetic and biological characteristics.It indicated that the PPMV aYQ strain possess satisfied immunogenicity,and can be used as the preferred pigeon paramyxovirus inactivated vaccine candidate strain.

关 键 词：鸽副黏病毒 疫苗 毒种 生物学特性 遗传稳定性 

分 类 号：S852.659.5[农业科学—基础兽医学] S858.39[农业科学—兽医学]