邯郸市猫疱疹病毒HD1株的分离与鉴定  

Isolation and identification of feline herpesvirus HD1 in Handan City

作　　者：余紫葳 蒋艳妹 宋金祥 崔德福 刘永相 范春艳 YU Ziwei;JIANG Yanmei;SONG Jinxiang;CUI Defu;LIU Yongxiang;FAN Chunyan(School of Life Science and Food Engineering,Hebei University of Engineering,Handan 056038,China;Hebei Leshou Duck Co.,Ltd.,Cangzhou 062250,China)

机构地区：[1]河北工程大学生命科学与食品工程学院,河北邯郸056038 [2]河北乐寿鸭业有限责任公司,河北沧州062250

出　　处：《畜牧与兽医》2024年第7期58-64,共7页Animal Husbandry & Veterinary Medicine

基　　金：河北省自然科学基金项目(C2021402005)。

摘　　要：为了探究猫疱疹病毒(FHV)在邯郸地区的分子流行病学特征,从邯郸地区不同动物医院收集到有呼吸道症状猫的眼、鼻分泌物共55份,经PCR初检后,将阳性样本接种于猫肾细胞(CRFK),细胞产生病变,经过蚀斑纯化、分子生物学检测、血清学鉴定、病毒形态鉴定,得到1株FHV。用gD基因引物扩增FHV分离株基因后能够得到特异性条带,且经过3轮蚀斑纯化后,得到1株亚克隆毒株,命名为FHV HD1。FHV HD1 P3代在CRFK上的病毒滴度为10^(6) TCID_(50)/100μL,并检测生长过程中8个时间点的病毒滴度,绘制分离株生长曲线。使用FHV-1特异性抗体进行间接免疫荧光检测,结果FHV HD1能够出现特异性荧光。BLAST分析表明FHV HD1株gD基因与其他FHV-1相似性达98.25%~99.91%。进化树分析表明与猫科动物源FHV-1亲缘关系较近,处于同一大分支,与犬疱疹病毒和其他动物疱疹病毒亲缘关系较远。本研究结果为FHV的进化分析和现阶段疫苗研发奠定了基础,对猫病毒性鼻气管炎的病原学、免疫学、临床诊断及分子生物学的研究具有重要参考价值。In order to explore the epidemic characteristics of feline herpesvirus in the Handan area,a total of 55 samples of eye and nose secretions of cats with respiratory symptoms were collected from different animal hospitals in the area.Initial PCR detection was performed,and positive samples were obtained and were inoculated on crandell reese feline kidney(CRFK)cells which showed pathological changes.Next,plaque purification,molecular biological detection,serological identification,and virus morphological identification were conducted,the positive samples were inoculated into the CRFK cells,and a strain of feline herpes virus(FHV-1)was obtained.Finally,specific bands were obtained by amplifying the FHV-1 gene using the gD gene primer,after three rounds of purification,a subclonal strain named FHV HD1 was obtained.The results showed that the virus titer of FHV HD1 P3 on CRFK was 10^(6) TCID_(50)/100μL.Virus titers were measured at 8 timing points during the growth,and the growth curve of the isolates was obtained.FHV-1 specific antibody was used for indirect immunofluorescence detection,and FHV-HD1 showed specific fluorescence.BLAST analysis showed that the gD gene of the FHV-HD1 strain was of 98.25%-99.91%similarity to the other FHV-1 strains.Phylogenetic tree analysis showed that it was closely related to feline source FHV-1 in the same clade,but was far related to canine herpesvirus and other animal herpesvirus.These results indicatd that the FHV-1 strain in the Handan area was evolutionarily stable at present,which laid a theoretical foundation for the evolutionary analysis of feline herpesvirus and the development of vaccine at the present stage.The results also possessedimportant reference value for the study of etiology,immunology,clinical diagnosis and molecular biology of feline viral rhinotracheitis.

关 键 词：猫疱疹病毒1型 分离鉴定 生长动力学 GD基因 

分 类 号：S852.615[农业科学—基础兽医学]