两株猪伪狂犬病病毒变异株的分离鉴定及gB、gC和gE基因的分子特征分析

doi:10.16431/j.cnki.1671-7236.2019.03.029

Abstract

Abstract:

To investigate the genetic variation of pseudorabies virus (PRV) in Henan province in recent years,in this study,the virus isolation and identification were performed by cell blind transmission,indirect immunofluorescence,Western blotting,plaque purification and transmission electron microscopy in brain tissue samples from suspected pseudorabies farms in Luohe and Zhongmou in 2017.The viruses titer and growth curve of the isolated strains were determined by 50% tissue culture infective dose (TCID₅₀),and the lethality of the mice was determined by mouse infection test.Futhermore,the gB,gC and gE genes were aligned and analyzed.Two strains of PRV were successfully isolated and identified by indirect immunofluorescence,plaque purification and transmission electron microscopy,named as HeN-LH strain and HeN-YM strain,respectively.The titers of HeN-LH and HeN-YM strains reached 10^8.35 and 10^6.63 TCID₅₀/mL at 36 h post infection on PK-15 cells.The lethality of the mice showed that the median lethal dose (LD₅₀) of HeN-LH and HeN-YM strains was 10^2.13 and 10^3.25 TCID₅₀,respectively.The evolutionary analysis displayed that the two isolates were closer to the PRV variants which were mainly epidemic in China since 2011.Amino acids sequences alignment showed that,similar to other variants,gB,gC and gE genes had multiple amino acid variations,and there were unique amino acid insertions and deletions at specific sites.In short,we successfully isolated and identified two PRV variants,and confirmed the current pseudorabies were mainly caused by the PRV variants,and the results could provide data for the prevention and control of pseudorabies and the selection of vaccine strains in Henan province.

Key words: pseudorabies virus (PRV); isolation and identification; plaque purification; infection test; sequence analysis

CLC Number:

S852.65

LI Xiang, GUO Zhenhua, RUAN Haiyu, QIAO Songlin, ZHANG Yifang, ZHANG Gaiping. Isolation and Identification of Two Pseudorabies Virus Variants and Analysis of Molecular Characteristics of gB,gC and gE Genes[J]. , 2019, 46(3): 881-890.

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Isolation and Identification of Two Pseudorabies Virus Variants and Analysis of Molecular Characteristics of gB,gC and gE Genes

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