畜牧兽医学报 ›› 2019, Vol. 50 ›› Issue (10): 2070-2078.doi: 10.11843/j.issn.0366-6964.2019.10.013

• 预防兽医 • 上一篇    下一篇

一株分离自免疫猪场的伪狂犬病病毒的鉴定与变异分析

王一鹏, 王亚文, 徐瑞涛, 林依丹, 李潭清, 宋勤叶*   

  1. 河北农业大学动物医学院, 河北省兽用生物制品工程技术研究中心, 保定 071000
  • 收稿日期:2019-04-01 出版日期:2019-10-23 发布日期:2019-10-23
  • 通讯作者: 宋勤叶,主要从事动物传染病与新型生物制品研究,E-mail:songqinye@126.com
  • 作者简介:王一鹏(1993-),男,河北廊坊人,硕士生,主要从事动物传染病与新型生物制品研究,Tel:0312-7528357,E-mail:15831625993@163.com
  • 基金资助:
    河北省科技计划项目(17226613D-2);河北农业大学科研发展基金(3003037)

Identification and Variation Analysis of Pseudorabies Virus from Vaccinated Pig Farm

WANG Yipeng, WANG Yawen, XU Ruitao, LIN Yidan, LI Tanqing, SONG Qinye*   

  1. Hebei Province Research Center of Veterinary Biologicals Engineering and Technology, College of Veterinary Medicine, Hebei Agricultural University, Baoding 071000, China
  • Received:2019-04-01 Online:2019-10-23 Published:2019-10-23

摘要: 为了明确河北某伪狂犬病疫苗免疫猪场的哺乳仔猪出现神经症状和死亡的病因,作者对发病仔猪进行研究时从脑组织中分离到1株病毒,通过PCR、免疫过氧化物酶单层试验(IPMA)、动物试验以及基因序列分析等试验,证明所分离的病毒为伪狂犬病病毒(PRV),命名为HBXT-2018株。皮下接种该病毒后24 h,家兔开始啃咬病毒接种部位皮肤,在44~68 h内全部死亡。抗PRV Bartha-K61株的血清对分离株的中和抗体效价低于1:8,而抗PRV变异株血清的中和抗体效价为1:58.9。与Bartha株比较,HBXT-2018株的gB和gC的核苷酸和氨基酸序列多处发生单个或连续几个碱基(或氨基酸)置换、插入和缺失突变,预测的gB和gC上的潜在抗原表位也发生了改变。基于gB、gC和TK的系统进化分析表明,HBXT-2018株与国内流行毒株尤其与2011年之后的流行毒株处于同一个进化分支,而与Bartha株及其他欧美毒株处于不同的进化分支。上述结果表明,分离的PRV HBXT-2018株为致病毒株,其与当前国内流行的PRV变异株具有相同的遗传特征,与Bartha株为代表的欧美流行株的遗传关系远。Bartha-K61株抗血清对分离株的中和作用不明显,提示gB与gC的突变可能与该毒株免疫逃逸、继而导致仔猪发病有关。

Abstract: In order to illuminate the cause for neurological sign and death in suckling piglets at a Bartha-K61-vaccinated pig farm in Hebei province, one wild stain of pseudorabies virus (PRV), named HBXT-2018, was isolated from the brain tissues of sick piglets and identified by PCR, Immunoperoxidase monolayer assay (IPMA), animal experiment and gene sequencing in this study. Experimental rabbits were subcutaneously injected with HBXT-2018 strain of 105TCID50, and they started to bite the inoculated sites at 24 h and all of rabbits died within 44-68 h. Neutralizing antibody titer of anti-PRV strain Bartha-K61 serum was lower than the dilution of 1:8, whereas that of anti-PRV variant serum was 1:58.9. Compared with Bartha strain, gB and gC nucleotide and amino acid sequences of HBXT-2018 had various mutations including single or continuous several base-pair or amino acid residue substitutions, insertion and deletion, and predicted antigenic epitopes changes happened in the amino acid sequences of gB and gC. Phylogenetic analysis based on gB, gC and TK showed that HBXT-2018 clustered in the same branch with those strains prevalent in China especially since 2011 whereas it was located at a different branch from Bartha strain and other PRVs in Europe and America. The results indicate that HBXT-2018 isolated in this study is a virulent strain with the same genetic characteristics as the PRV variants currently prevalent in China while it has a remarkably distant evolutionary relationship with Bartha strain. Anti-Bartha-K61 strain serum did not neutralize this isolate well, which suggests that the mutations in gB and gC might be related to the viral immune escape causing piglet pseudorabies in this pig farm.

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