猪伪狂犬病病毒鲁A株的分离鉴定

Isolation and Identification of LA Strain of Pseudorabies Virus

从山东省某些猪场疑似猪伪狂犬病发病仔猪的脑及内脏中分离到多株病毒 ,对其中一代表毒株进行了全面鉴定。该分离毒株在兔肾原代细胞 ( RK)上和鸡胚成纤维细胞 ( CEF)上连传 5代 ,均出现典型细胞病变 ,再接种于 RK- 13细胞、BHK- 2 1细胞、Vero细胞、PK15细胞和 14 3TK- 细胞 ,均出现典型细胞病变 ;在 RK- 13细胞上的 TCID50 为10 - 6 .52 / 0 .1m L ;在电镜下可见到典型的伪狂犬病病毒粒子 ;对氯仿、乙醚敏感 ,5 6℃ 30 m in灭活 ;能被伪狂犬病病毒标准阳性血清中和 ;病毒接种于家兔和小鼠 ,均出现典型伪狂犬病症状与病变 ;提取所分离病毒的 DNA作为模板 ,应用特异性引物 ,以 PCR方法可扩增出伪狂犬病病毒 g D基因中 2 72～ 5 34nt之间 2 6 2 bp长的特异性片段。上述结果证明 ,所分离病毒为猪伪狂犬病病毒 ,并将其命名为猪伪狂犬病病毒鲁

Several virus strains have been isolated from the samples of brains and visceral organs of death piglets from some farms in Shandong province and one of them was identified.The virus was continually inoculated 5 generations onto rabbit kidney cell and chicken embryonic fibroblast cell(CEF) and induced typical cytopathic effect(CPE).The typical CPE could be seen when culturing the virus on RK 13 cell,PK15 cell,Vero cell,BHK 21 cell and 143 TK - cell and the TCID 50 of the isolated virus was 10 -6 52 /0 1 mL and the typical Pseudorabies virus particles could be found under electronmicroscope.The isolated virus was sensitive to chloroform and ethylether and inactivated for 30 minutes at 56℃,and could be neutralized by the positive serum to Pseudorabies virus.The typical clinical syndrome could be observed when four rabbits and 10 mice were injected with the isolate.The sequence of 262 base pair lying in gD gene could be amplified with the DNA extracted from the isolate and a set of oligonucleotide primers.All of the evidences indicated that the isolated virus was Pseudorabies virus(PRV) and was named as LA strain of Pseudorabies virus.