伪狂犬病病毒Guizhou-DY株的分离鉴定及致病性研究

Identification and Pathogenicity Study of Pseudorabies Virus Guizhou-DY Strain

通过采集疑似猪伪狂犬病流产胎儿病料，采用细胞接毒分离培养、病毒蚀斑纯化、病毒的形态结构观察、易感动物接种试验和病毒核酸鉴定分离到1株伪狂犬病病毒（PRV），命名为PRVGuizhou—DY株（GenBank登录号为JX417716）；为了摸清其理化特性，进一步对Guizhou—DY分离株进行了耐热、耐酸及脂溶性敏感性试验、核酸类型鉴定及病毒毒价测定，同时进行病毒致病性试验。分离鉴定结果表明：病料接种Vero细胞盲传至第2代时产生明显CPE，盲传至第4代可出现稳定的CPE；病毒粒子在电镜下呈椭圆形、有囊膜、直径为120～180nm，核内可见呈晶格状排列的病毒包涵体；病毒悬液接种家兔能引起家兔奇痒、麻痹死亡；理化特性研究表明：该病毒是一种耐热不耐酸且对氯仿敏感的DNA病毒，gE全基因序列与PRVGZ—Z1株、Fa株及Ea株的相似性分别为97．60A、98．9％和99．4％，其TCID50为10-9.71·100μL-1，PFu为109·mL-1. 攻毒组猪体温升高，临床症状较为明显，在攻毒后第5天开始出现中和抗体，第21天达到最高。该研究可为PRV病原学研究提供参考。

One pseudorabies virus was isolated from aborted swine fetus of a hoggery of Guizhou province, and was identified. It was named as the PRV Guizhou-DY strain （GenBank accession number JX417716）. A lot of experiments had done in the study, e g heat resistance test, acid re- sistance test and fat-soluble sensitivity test, identification of nucleic acid type, and virus pathoge- nicity test. The inoculated Vero cells showed the typical cytopathogenic effect when cultured to the second generation, and the stable cytopathogenic effect occurred till the fourth generation. By transmission electron microscopy, the mature virions with envelope were round or ellipse with a diameter of 120 to 180 nm, and viral inclusion bodies arranged like crystal lattice were visible in cell nuclei. The rabbits injected with the isolated virus showed the typical clinical syndrome of pruritus, palsy, opisthotonos and death in the end. Experiments on the virus＇s physicochemical properties showed that PRV Guizhou-DY Strain was a heat-resistant but not acid and chloroform sensitive DNA virus. Sequence analysis of gE genome some variation compared with GZ-Z1 strain, Fa strain showed that PRV Guizhou-DY Strain had and Ea strain, the homology was 97.6%0, 98.9% and 99.4G. And its TCIDs0 was 10-9.71 100 μL-1 , PFU was 109 mL-1. The challenge group pigs developed a fever, and the clinical symptoms are more obvious, neutralizing antibodies began to appear at the 5th day post infection and at its peak at the 21st day post infection. This study can provide reference for the study of the viral etiology.