【作者】 戴秀美； 王敏； 王蕾； 郭树源； 常维山；

【Author】 Dai Xiumei;Wang Min;Wang Lei;Guo Shu-yuan;Chang Weishan;Key Laboratory of Animal Biotechnology and Disease Control and Prevention of Shandong Province,College of Veterinary Medicine,Shandong Agricultural University;

【机构】 山东农业大学动物科技学院；

【摘要】 应用Vero细胞从山东省诸城市疑似犬瘟热感染貉的肝脏、脾脏、肺脏等组织病料的研磨上清液中分离出1株病毒。分离株经Vero细胞传至第4代出现典型的细胞病变效应,经毒力测定、血清学、RT-PCR检测及基因测序、回归动物试验证实为犬瘟热病毒,命名为ZC株。采用RT-PCR方法分段克隆分离株的全基因组序列并测序,依次拼接得到全基因组序列进行序列比对分析。结果表明,CDV-ZC株全基因(GenBank登录号KJ994343)与美洲基因型犬瘟热强毒株A75/17核苷酸同源性高达96.5%,而与疫苗株Onderstepoort、CDV3等亲缘关系较远,同源性为91.6%-92.0%。血凝素基因序列分析表明,CDV-ZC株与国内野毒株LN(13)2、GP株等以及日本野毒株UENO、HAMA等共同归属于Asia-1基因型,在H蛋白信号淋巴细胞激活因子受体结合区的542-544位增加了1个潜在N-连接糖基化位点。致病性强毒株CDV-ZC的成功分离及全基因序列测定加深了我们对当前中国CDV流行株遗传变异情况的了解,对CD的有效预防、诊断及控制具有指导意义。更多还原

【Abstract】 A virus strain was isolated by Vero cells from supernatant of cell extracts of liver,spleen,lung that collected from a raccoon dog with suspected canine distemper virus(CDV) infection in Zhucheng City,Shandong Province,China.Typical cytopathic effect(CPE) was observed in Vero cells after four passages.It identified as CDV by virulence investigation,serology neutralization test,RT-PCR and animal regression experiment and named CDV Zhucheng(CDV-ZC) strain.Full-length genome of strain CDV-ZC was amplified,sequenced and aligned with other CDV strains.The complete genome nucleotide sequence of CDV-ZC(Accession number KJ994343)exhibited highly homology to the standard virulent strain A75/17(96.5%at nt level),while identity to the vaccine strains ranged from 91.6%(for Onderstepoort) to 92.0%(for Phoca/Caspian).Phylogenetic analysis based on the hemagglutinin(H) gene demonstrated that CDV-ZC strain belonged to Asia-1 genotype,along with Chinese wildtype strain LN(13)2,GP and Japanese strain UENO,HAMA and so on.Inspecting in the H protein alignment,a novel potential N-linked glycosylation sites(N-X-S/T) was observed at position 542-544,which located at H protein receptor binding region.The successful isolation and complete sequences determination of highly pathogenic and virulent strain CDV-ZC will provide a better understanding of the genetic variations of CDVs in China and is likely to be helpful to guide efficient preventive,diagnostic,and control strategies for CD.更多还原