猫疱疹病毒1型的分离鉴定及生物学特性分析  

Isolation,Identification and Biological Characteristics of Feline Herpesvirus⁃1

作　　者：陶伟杰 刘雪娇 宋晓莉 程宝钰 于永乐 单虎 张传美 TAO Weijie;LIU Xuejiao;SONG Xiaoli;CHENG Baoyu;YU Yongle;SHAN Hu;ZHANG Chuanmei(College of Veterinary Medicine,Qingdao Agricultural University,Qingdao 266109,China)

机构地区：[1]青岛农业大学动物医学院,青岛266109

出　　处：《病毒学报》2023年第4期1053-1061,共9页Chinese Journal of Virology

基　　金：2021年山东省专业学位研究生教学案例库(项目号:SDYAL21174),题目:动物疫病分子诊断技术案例库。

摘　　要：为了丰富对猫疱疹病毒1型(Feline herpesvirus⁃1,FHV⁃1)基因组变异情况的研究,并为疫苗研发提供候选毒株,本研究对250份猫眼鼻拭子和肺组织样品进行PCR检测,对FHV⁃1阳性样品进行氨基酸变异和系统发育分析,并进行毒株的分离鉴定及动物回归试验。结果显示,250份临床样品中FHV⁃1型阳性率为20.8%(52/250)。测序得到10个毒株的8个囊膜蛋白基因序列,氨基酸序列分析显示9个毒株的gI基因发生非同义突变(M165T),QN⁃1毒株gD基因发生非同义突变(A342T)。系统发育分析表明毒株间变异小、进化距离短。病原分离鉴定获得1株FHV⁃1毒株(命名为FHV/BJ⁃1),病毒滴度为10^(6.65) TCID_(50)/0.1 mL。动物回归试验结果显示,接毒组家猫在8 d内全部死亡,剖检及HE染色发现肺部病变严重,有淤血及炎性细胞浸润,气管缺少上皮细胞,且鼻组织、气管和肺组织的病毒载量最高。上述结果表明,FHV⁃1流行毒株基因保守性好,各毒株之间主要囊膜蛋白变异程度低。分离株FHV/BJ⁃1毒株为强毒株,是疫苗评价的理想毒株,为疫苗候选株的筛选和疫苗的研发奠定了基础。In order to enrich the research on the variability of feline herpesvirus⁃1(FHV⁃1)genome and provide candidate strains for vaccine research and development,250 cat eye and nose swabs and lung tissue samples were detected by PCR.Amino acid variation analysis and phylogenetic analysis were performed.Isolation and identification of virus strains and animal regression experiments were carried out.Results showed that the positive rate of FHV⁃1 in 250 clinical samples was 20.8%(52/250).8 envelope protein gene sequences of 10 strains were obtained by sequencing,and amino acid sequence analysis showed that the gI gene of 9 strains showed non⁃synonymous mutation(M165T),and the gD gene of QN⁃1 strain showed non⁃synonymous mutation(A342T).Phylogenetic analysis showed that variation among strains was small and the evolutionary distance was short.A strain of FHV⁃1(named FHV/BJ⁃1)was isolated and identified with 10^(6.65) TCID_(50)/0.1 mL.The results of animal regression experiment showed that all cats in the infected group were died within 8 days.Animal autopsy and HE staining showed severe lung lesions with congestion and inflammatory cell infiltration,and lack of epithelial cells in the trachea.The results of virus tissue distribution showed that nasal tissue,trachea and lung tissue had the highest viral load.The above results showed that the genes of the epidemic strains were well conserved,and the variation degree of the main envelope proteins among the strains was low.The isolate FHV/BJ⁃1 is a high virulent strain,which is ideal for vaccine evaluation.This study lays a foundation for screening vaccine candidate strains and the research and development of vaccines.

关 键 词：猫疱疹病毒1型 序列分析 分离鉴定 动物回归试验 

分 类 号：S855.3[农业科学—临床兽医学]