环洞庭湖地区活禽市场鸭源 NDV 的分离鉴定及遗传演化分析

摘要

A Newcastle disease virus,named duck/hunan/yy 858/14,was isolated from 14 samples of the healthy ducks in live poultry markets around Dongting Lake region.It was identified with hemagglutina-tion test,hemagglutination inhibition test and RT-PCR.The virus F gene was amplified by RT-PCR,se-quenced and analyzed by DNA Star and Lasergene software.The phylogenetic tree of F gene was construc-ted .The result showed that the total length of the F gene was 1 792 bp,ORF was 1 662 bp.The F gene encodes 553 amino acids.The virus named duck/hunan/yy 858/14 belong to genotype 3 of Class I virus. It′s composition of amino acid cleavage site was 112 ERQERL117 ,which was typical molecular characteristic of Newcastle disease virus.Homology analysis showed that the homology was low,73.4% and 71.6% re-spectively among duck/hunan/yy 858/14,V4 and La Sota vaccine strains in ClassⅡ.The homology was high,91.2% and 91.0% respectively among duck/hunan/yy 858/14,Chicken/HongKong/1525.16/2005 and Duck/China/08-004/2008,there is closer genetic relationship to wild-bird Jilin strain J36/13.%从洞庭湖地区周边县市几个活禽市场的健康家鸭中分离到14株具有血凝性的病毒，通过血凝、血凝抑制试验及 RT-PCR 鉴定，其中1株为新城疫病毒，命名为 Duck／hunan／yy858／14。通过对该病毒的 F基因全长扩增，并转化、克隆、测序及构建 F 基因的遗传进化树，推导其核苷酸序列。序列测定及遗传进化分析显示，该毒株 F 基因全长为1792 bp，ORF 为1662 bp，可编码553个氨基酸，其氨基酸裂解位点的组成为112 ERQERL117，具有典型新城疫病毒弱毒的分子特征，与我国内地首次分离到的 Class I 毒株 NDV08-004、吉林野鸟毒株 J36／13、香港活禽市场分离株 Chicken／HongKong／1525．16／2005在同一分支，属 Class I类基因3型。同源性分析显示该毒株与 ClassⅡ中弱毒疫苗株 V4和 La Sota 的核苷酸及氨基酸的同源性均较低，分别为73．4％和71．6％；与香港活禽市场分离株 Chicken／HongKong／1525．16／2005和首例内地分离到的鸭源新城疫病毒毒株 Duck／China／08-004／2008的核苷酸同源性分别为91．2％、91．0％；与吉林野鸟毒株J36／13的核苷酸同源性最高为98．3％，推测与野鸟毒株有共同来源。