一株新疆野鸟源新城疫病毒的分离鉴定、F基因克隆及遗传进化分析

Isolation and Identification,F Gene Cloning and Evolutionary Analysis of Newcastle Disease Virus Isolated from Wild Birds in Xinjiang

为了解新疆野鸟新城疫病毒(Newcastle disease virus,NDV)感染情况,分析其分子特征和基因变异特点,防止新城疫的暴发和蔓延,本试验用9日龄SPF鸡胚进行病毒的分离传代,HA、HI试验和RT-PCR方法对位于"东非—西亚迁徙线"福海县的野鸟进行了NDV检测,分离到1株野鸟源NDV,并命名为NDV/Pintail/CH(XJ)/01/2016。结果表明,该NDV分离株F基因ORF长1 662bp,编码553个氨基酸,F基因碱性裂解位点序列为^(112)G-R-Q-G-R-L^(117),符合弱毒株特征;遗传进化分析显示,NDV/Pintail/CH(XJ)/01/2016与乌克兰鸽源分离株Doneck/3/968、比利时鸭源分离株Simeonovgrad核苷酸同源性均为99.7%,属于NDV ClassⅡ基因Ⅱ型。而上述3株病毒均与疫苗毒La Sota具有较高同源性(99.5%~99.8%)。本研究结果为家禽NDV外流进入自然环境提供了论据。

In order to understand inflection of Newcastle disease virus（NDV）of Xinjiang wild birds,through analysis of molecular characteristics and genetic variation,to prevent the outbreak and spread of Newcastle disease,isolation of the virus was performed in 9days old specific pathogen free（SPF）chicken embryos,detected the wild birds in Fuhai county which was located in the migration line of East Africa-West Asia by HA,HI test and RT-PCR method.And then 1strainof NDV from wild birds was isolated,named as NDV/Pintail/CH（XJ）/01/2016.As the result,ORF of Fgene from the NDV isolate was 1 662 bp,encoding 553 amino acids.The motif of the cleavage site was ^（112）G-R-Q-G-R-L^（117）,which was consistent with the characteristics of avirulent NDV strains.The phylogenetic analysis showed that NDV/Pintail/CH（XJ）/01/2016 nucleotide sequence homologies of F gene between reference strain Doneck/3/968 and mule Simeonovgrad strain homology were high and reached 99.7%,it belonged to genotypeⅡ of ClassⅡ NDV.All of the three viruses had high homology to the vaccine strain La Sota,which were 99.5% to99.8%.According to this research conclusion we provided evidence that poultry NDV outflowed into the natural environment.