一例猪伪狂犬病病毒的分离鉴定

从临床疑似猪伪狂犬病发病仔猪的脑组织等病料中,经PCR扩增出大小为217 bp的伪狂犬病病毒gp50的基因片段,结果证实为猪伪狂犬病病毒(porcine pseudorabies virus,PRV)感染。随后采用BHK-21细胞进行猪伪狂犬病病毒的分离培养,该分离株经细胞传代培养5代后,能够产生典型的细胞病变,经PCR鉴定为伪狂犬病病毒,其病毒感染力达10^8.68 TCID₅₀/0.1mL。最后用10^7.0 TCID₅₀/mL病毒培养物接种家兔,48 h后注射部位出现典型瘙痒、皮肤破损等症状,于72 h后全部死亡。结果表明,该伪狂犬病病毒分离株对易感动物具有高致病性,为进一步开展该病毒流行病学、致病机理、疫苗免疫及诊断研究奠定了基础。

The Isolation and Identification of the Porcine Pseudorabies Virus

The brain tissue of piglet was obtained and the 217 bp long fragment in gp50 gene was amplified by the polymerase chain reaction (PCR). Therefore, the piglet was confirmed as porcine pseudorabies virus infection. Subsequently, the porcine pseudorabies virus was isolated on BHK-21 cells after 5 generations. The cells displayed typical cell lesions and were identified as porcine pseudorabies virus by PCR. The virus infectivety of the pseudorabies virus was 10^8.68 TCID₅₀/0.1 mL by Reed-Muench. Finally, the rabbits were inoculated with 10^7.0 TCID₅₀/mL virus. On 48 h the injection sites showed typical itching, skin damage and other symptoms. The rabbits were dead after 72 h by the inoculation of pseudorabies virus. Consequently, the results indicated that the isolated pseudorabies virus was highly pathogenicity to susceptible animals. It would be the research foundation of the virus epidemiology, pathogenesis,vaccine and diagnositic.