猪伪狂犬病病毒GXBB株的分离鉴定及gE基因的克隆分析

Isolation and Identification of GXBB Strain of Porcine Pseudorabies Virus and Sequence Analysis of gE Gene

从广西玉林博自某猪场采集的发病仔猪大脑和内脏病料中分离到一株病毒。病毒接种家兔后引起典型的奇痒、神经症状，接种PK-15细胞出现典型的细胞病变，病毒效价（TCID50）为10^-7.22／0．1ml。设计扩增PRVgE胞外区基因的引物，能扩增出约947bp的特异性片段，将扩增出的目的片段进行克隆、测序，并与国内外不同PRV毒株进行分析比较，发现该毒株与国内MinA株、Ea株、SH株、LA株、GXB株、GXW株核苷酸同源性在98．7％～99．4％之间，氨基酸同源性在98．1％～99．1％之间，上述结果证实该分离毒株为伪狂犬病病毒，命名为GXBB株。GXBB株与国内流行毒株同源性很高，说明目前广西PRV流行株变异不大。这为下一步广西伪狂犬病的预防和净化工作提供科学的理论基础。

One virus was isolated from brain and visceral organs of diseased piglets from a pig farm in Bobai of Yulin of Guangxi. The virus that infected rabbits caused typical pseudorabies clinical symptom, induced typical cytopathogenic effect （CPE） after inoculated in PK-15 cell. And virus TCID50 was 10^-722/0.1 ml. A specific band about 947 bp was obtained by PCR amplification with a pair of primers designed according to a reference. The PCR product was cloned , sequenced and compared with MinA strain,Ea strain,SH strain,LA strain,GXB strain,GXW strain, the result displayed 98.7% to 99.4% nucleotide sequence homology and 98.1 % to 99. 1% amino acid sequence homology, so it was comfirmed as pseudorabies virus and it was named GXBB strain. Phylogenetic analysis revealed that the PRV GXBB strain and other PRV strains were closely related and there were few mutations about the epidemic PRV strains in Guangxi at present. The study provides the scientific theoretical basis for prevention and eradication of PRV.