摘要
针对我国东北地区获得高免新城疫(NewcastleDisease,ND)抗体鸡群仍然发生新城疫(ND)的情况,应用分子流行病学技术,对所分离的18株鸡源新城疫病毒(NewcastleDiseaseVirus,NDV)和1株鸽源NDV进行系统发育进化分析。将NDV分离株通过差速离心纯化,以SDS-蛋白酶K(或Trizol法)提取病毒RNA,RT-PCR扩增其融合蛋白(F)基因532bp或280bp关键片断,经回收克隆到pMD18-T载体上进行核苷酸序列测定(GenBank序列号:AY208680-AY208698)。核苷酸和氨基酸序列进行同源性比较,结果表明各毒株之间核苷酸同源性为83.1%～100%,氨基酸同源性为85.5%～100%;系统发育进化树分析显示,其中JL-12、HLJ-3为弱毒株,与疫苗株V4同属于基因Ⅰ型;其余17株均为强毒株,其中HLJ-4、JL-14为Ⅵ型,其余15株均为基因Ⅶ型,由此可见东北地区目前新城疫的流行是由3种不同基因型的NDV毒珠(Ⅰ型、Ⅵ型、Ⅶ型)所发,但以基因Ⅶ型为主,这与我国其他地区和世界上其他国家ND的流行情况趋于一致。
In recent years, there occurred non-typical ND in vaccinated chicken flock of northeast china. To have insight into the molecular epidemiology of Newcastle disease virus （NDV） prevailed. 18 NDV strains isolated from chicken and 1 NDV isolated from pigeon using SPF chicken embryos were studied with RT-PCR and subsequence analysis of the region of 255 nucleotides covering the fusion （F） protein cleavage site （GenBank number： AY208680- AY208698）. JL-12,HLJ-3 have the amino acid sequence 112GR （K）QGRL117 at the F2/F1 cleavage site （112-117） of F protein, which confirmed that they were lentogenic, the other isolated NDV strains have 112RRQR （K）RF117,which confirmed that they were velogenic. The genetic similarities among the viruses isolated are 83.1%-100%, protein similarities among the viruses are 85.5%-100%. Phylogenetic tree based on 255 bp （position 338-592 nO region of F gene was established. The tree indicated JL-12,HLJ-3 were of genotype Ⅰ the same as V4; HLJ-4, JL-14 were of genotype Ⅵ. Other 15 strains were grouped under genotype Ⅶ. All the results indicated that NDV in northeast china is very complex, Both old genotype Ⅰ （2/19）and Ⅵ （2/19）and novel Ⅶ （15/19）had been isolated and genotype Ⅶ were most prevailing strains.
出处
《畜禽业》
2006年第1期26-30,共5页
Livestock and Poultry Industry
关键词
NDV
F基因
分子流行病学
系统发育进化树
Newcastle Disease Virus
F gene
Molecular Epidcmiology
Phylogenetic Tree
