一例伪狂犬病毒野毒株感染病例的病原学诊断

Etiological diagnosis of a case of pseudorabies virus wild strain infection

对一例疑似伪狂犬病毒(PRV)感染病例进行实验室检测和病原的分离培养。采集病猪的脏器,进行PRV gE基因的PCR检测,病料接种非洲绿猴肾细胞(Vero)并传代培养,观察细胞病变效应(CPE),PCR监测第3代、第5代和第7代培养物PRV的gE和gG基因核酸,采用免疫过氧化物酶单层细胞染色法(IPMA)检测病毒在Vero细胞中的增殖动态。结果表明,病猪心脏、肺脏和脾脏为PRV gE基因核酸阳性,Vero细胞在接种病毒48 h,变圆、融合;第3代、第5代和第7代培养物中gE和gG基因核酸均为阳性,第7代培养物与PRV阳性血清呈特异性反应,病毒感染后第6 h、Vero细胞中即可检测PRV抗原,第24 h绝大部分细胞呈PRV抗原阳性。该病例为PRV野毒株感染,所分离的PRV毒株对Vero细胞适应力强。

In this study,a suspected case of pseudorabies virus(PRV)infection was tested in laboratory and the pathogen was isolated and cultured.The gE gene of PRV was detected by polymerase chain reaction(PCR)from the organs of infected pigs.The samples were inoculated with Vero kidney cells and cultured to observe the cytopathy effect(CPE).The gE gene and gG gene nucleic acid of the 3rd,5th and 7th generation cultures of PRV were monitored by PCR.The proliferation of the virus in Vero cells was detected by immunoperoxidase monolayer staining(IPMA).PCR results showed that PRV gE gene nucleic acid was positive in the heart,lung and spleen of infected pigs,and Vero cells became round and fused 48 h after inoculation.The gE and gG gene nucleic acid in the 3rd,5th and 7th generation culture were positive,and the 7th generation culture had a specific reaction with PRV positive serum.PRV antigen could be detected in Vero cells as early as 6 h after virus infection,and most of the cells were PRV antigen positive at 24 h after virus infection.This case was infected by a wild PRV strain,and the isolated PRV strain was highly adaptable to Vero cells.