猪伪狂犬病病毒LC株的分离鉴定及其重要功能基因的进化分析

为确诊广东某猪场母猪流产发病原因，本研究收集该猪场流产死胎的脑、淋巴结、肺脏混合液，进行PCR鉴定、病毒分离培养、半数组织培养感染剂量（tissue culture infective dose，TCID₅₀）测定、猪伪狂犬病病毒（pseudorabies virus，PRV）重要功能基因（gB、gC、gD、gE）序列测定和进化分析及动物回归试验。结果显示，病料混合液为PRV阳性，接种Vero细胞传至第3代即出现稳定的细胞病变（CPE），第5代TCID₅₀达到10^-6.8/0.1 mL，PRV gB、gC、gD、gE基因序列测定、同源性及进化树分析显示为，PRV中国变异株，命名为LC株。动物试验显示，LC株对12周龄猪具有一定致病性，可形成PRV典型临床症状及病理变化。本研究分离到一株PRV流行毒株，推测当前使用疫苗Bartha-K61株尚无法完全控制新毒株的流行。

Isolation,Identification and Phylogenetic Analysis of Important Functional Genes of Porcine Pseudorabies Virus LC Strain

To find out the reason of the reproductive failure in pregnant sows in a hoggery in Guangdong, the mixture with brain, lymph nodes and lungs of farm abortion stillbirth were identified by PCR, virus isolation and culture, tissue culture infective dose (TCID₅₀) assay, homology and phylogenetic analysis of the important functional genes (gB, gC, gD and gE) and animal test. The results showed that the mixture was proved to be porcine pseudorabies virus (PRV) positive samples. The typical cytopathogenic effect was induced in the third passage of Vero cell and the titer of the fifth passage was 10^-6.8/0.1 mL. The sequence analysis and phylogenetic relationship of gB, gC, gD and gE genes showed that it was a Chinese PRV variant, which was named as LC strain. The typical pseudorabies clinical and pathological symptoms were presented in 12-week-old piglets inoculated with LC strain. The results demonstrated that a local pseudorabies virus had been isolated, suggesting that the Bartha-K61 vaccine was not fully effective for controlling the current epidemic of pseudorabies in China.