猪伪狂犬病病毒的分离鉴定及其gE基因序列分析

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

从福建省某猪场疑似伪狂犬病的发病3日龄仔猪的脑、肺脏中分离到1株病毒。该病毒接种家兔出现了典型的伪狂犬病症状,接种PK-15细胞36h后出现了圆缩、集聚、脱落等典型的细胞病变,猪伪狂犬病病毒阳性血清能特异性中和该分离病毒。根据已发表的伪狂犬病病毒(PRV)gE基因的序列,设计并合成了一对引物,采用PCR方法可扩增特异性298bp的DNA片段。测序结果与GenBank中有代表性的6株参考毒株相应基因序列比较,核苷酸和氨基酸序列同源性分别为95.5%～99%和91.8%～98%。系统进化树结果表明分离株与湖北分离株Ea株亲缘关系最近。

One local strain of Porcine pseudorabies virus was isolated from the brain of a 3-day old pig from Fujian Province. Rabbits were inoculated with the pseudorabies virus isolate showed typical PRV symptoms.The isolated virus could cause typical cytopathogenic effect （CPE） in PK-15 cell at hour 36 after infection such symptoms as shrunk, aggregated and detached. The isolated virus could be neutralized with the positive sera. The DNA extracted from the virus isolate was subjected to polymerase chain reaction （PCR） analysis, a set of primer which can be used to amplify 298 bp fragment in the gene of PRV was designed. The gene was sequenced and compared with those of six other PRV strain in GenBank.The results showed that the gE gene of PRV shared 95.5%- 99% nucleotide homology and 91.8%-98% amino acid homology with strains.