新城疫病毒分离株的生物学特性鉴定及F蛋白基因序列分析  

Biological Characteristics and Sequence Analysis of Fusion Genes of Newcastle Disease Virus Isolates

作　　者：姚春峰[1] 刘文博[1] 胡顺林[1] 马怀良[1] 薛峰[1] 刘慧谋[1] 刘秀梵[1] 

机构地区：[1]扬州大学农业部畜禽传染病学重点开放实验室,江苏扬州225009

出　　处：《病毒学报》2009年第2期117-124,共8页Chinese Journal of Virology

基　　金："十一五"国家科技支撑计划(2006BAD06A03);国家自然科学基金重点项目(30630048);江苏省教育厅指导计划项目(06KJD230211)

摘　　要：通过部分生物学特性鉴定、RT-PCR及F基因的序列测定与遗传进化分析,对2005~2006年从我国江苏省和广西省部分地区的发病鸡群和鹅群中分离到的20株新城疫病毒(NDV)进行了研究。各分离株经典毒力测定结果显示:MDT在45.3h^58.2h之间,ICPI在1.61~2.00之间,均为新城疫病毒强毒株特征。血凝解脱及血凝素热稳定性试验显示:各分离株的血凝解脱时间短,血凝素热稳定性较差,符合NDV强毒株的特征。F基因的序列测定表明,分离株之间的核苷酸序列具有79.7%~100%的同源性,与疫苗株LaSota的同源性为78.1%~83.4%;与国内标准强毒株F48E8同源性为80.2%~90.1%。推导其氨基酸序列分析表明,各分离株的F蛋白的裂解位点氨基酸组成为112R-R-Q-R/K-R-F117,具有NDV强毒株特征,与毒力测定结果相符。根据序列所绘制系统进化发生树,表明20株NDV分离株中有18株为基因Ⅶd型,2株为基因Ⅲ型。Twenty Newcastle disease virus （NDV） strains were isolated from chickens and geese in the field outbreaks during 2005 and 2006 in some regions of Jiangsu and Guangxi province. Assessment of the virulence by MDT and ICPI, RT-PCR and sequence analysis of fusion protein gene were used to compare the properties of NDV isolates. The results indicated that MDT and ICPI of the isolates were 45.3h-58. 2h and 1.61-2.00 respectively, which confirmed that the all NDV isolates were highly virulent. And their hemagglutinin were not resistant to heat and belonged to fast pattern of elution. The results of nucleotide sequencing and phylogentic analysis of fusion protein gene showed that the twenty strains shared homology from 79.7% to 100% among themselves, from 78.1% to 83.4% and from 80.2% to 90.1% with NDV LaSota, F48E8, respectively. The putative amino acid sequences of fusion protein at the cleavage sites of all the isolates were 112R-R-Q-R/K-R-F117, with the motif characteristics of the virulent NDV strain, which was in accordant with the results of assessment of the pathogenicity. The phylogentic tree based on sequences of fusion protein gene variable regions （47-420nt） revealed that the 18 strains belonged to subgenotype Ⅶd and the others belonged to an old genotype Ⅲ of NDV, revealing that subgenotype Ⅶd virus was responsible for the NDV outbreaks in some regions of Jiangsu and Guangxi promince recently.

关 键 词：新城疫病毒 生物学特性 毒力 F基因 序列分析 

分 类 号：S858.31[农业科学—临床兽医学]