中国畜牧兽医

• 疾病防治 • 上一篇    下一篇

猪伪狂犬病病毒的分离鉴定及免疫原性初步研究

尹秀凤,丁美娟,张海涛,秦进进,孙?,许秀梅,汪爱芬,张小飞,薛家宾   

  1. (南京天邦生物科技有限公司,江苏南京 211102)
  • 收稿日期:2013-10-08 出版日期:2014-04-20 发布日期:2014-05-27
  • 通讯作者: 张小飞。E-mail:xiaofei0804@sina.com
  • 作者简介:尹秀凤(1975—),女,山东人,硕士,研究方向:动物传染病预防。
  • 基金资助:

    江苏省科技成果转化专项资金项目(BA2010021)。

Isolation and Identification of Porcine Pseudorabies Virus ZJ Strain and Research of its Immunogenicity

YIN Xiu-feng,DING Mei-juan,ZHANG Hai-tao,QIN Jin-jin,SUN Yan,XU Xiu-mei,WANG Ai-fen,ZHANG Xiao-fei,XUE Jia-bin   

  1. (Nanjing Tianbang Bio-industry Co.,Ltd.,Nanjing 211102,China)
  • Received:2013-10-08 Online:2014-04-20 Published:2014-05-27

摘要: 采集浙江某猪场疑似猪伪狂犬病发病仔猪的脑、脾脏等组织病料,经PCR检测为猪伪狂犬病病毒(pseudorabies virus,PRV)野毒感染,用BHK-21细胞进行病毒的分离培养,结果显示该病毒能引起典型的细胞病变,第4代病毒液毒价达107.0 TCID50/mL;PCR和动物回归试验结果表明该分离株为PRV,并将其命名为PRV ZJ株。将第4代病毒液制备成油乳剂灭活苗,免疫2 kg左右家兔,免疫后28 d采血并攻毒,测定其免疫原性,结果显示血清中和指数为13490,保护率为100%。本试验结果表明PRV ZJ株有很好的免疫原性,为进一步开展疫苗研究奠定基础。

关键词: 猪伪狂犬病病毒; 分离鉴定; 灭活疫苗; 免疫原性

Abstract: The material from the clinical samples of cerebrum and spleen in the pigs characterized by pseudorabies, proved to be porcine pseudorabies virus (PRV) positive samples through the amplification of gE gene of PRV, was inoculated into the BHK-21 cells and serially passaged 4 times. The isolate could induce typical cytopathogenic effect in cells and the titer of the fourth passage was 107.0 TCID50/mL. Identification was done by PCR method and four rabbits were injected with the fourth generation cell cultures of the isolate, and the specific band could be amplified and the typical clinical syndrome could be observed. It was indicated that the isolated virus named ZJ strain was PRV. The virulent PRV ZJ strain was killed with formalin and used as vaccine antigen with oil adjuvant. The rabbits were injected with 1 mL vaccine, 28 days after inoculation, serum antibodies were detected and healthy rabbits were challenged by PRV ZJ virulent strain, and the protection rate and serum neutralization index of antibodies were tested. The results showed that protection rate and serum neutralization index in rabbits were 100% and 13490,respectively.This study showed that PRV ZJ strain had good immunogenicity, and provided a basis for further research on vaccination of PRV.

Key words: porcine pseudorabies virus; isolation and identification; inactivated vaccines; immunogenicity