畜牧兽医学报 ›› 2015, Vol. 46 ›› Issue (5): 815-823.doi: 10.11843/j.issn.0366-6964.2015.05.018

• 预防兽医 • 上一篇    下一篇

一株重组鸡传染性支气管炎病毒的分离及结构蛋白基因变异和血清型分析

吴翠兰,何怡宁,李和鸣,孙新宽,谭渝才,韦天超,磨美兰*,韦平*   

  1. (广西大学动物科学技术学院,南宁 530005)
  • 收稿日期:2014-09-10 出版日期:2015-05-23 发布日期:2015-05-23
  • 通讯作者: 磨美兰,E-mail:momeilan@163.com;韦平,E-mail:pingwei8@126.com
  • 作者简介:吴翠兰(1988-),女,苗族,湖南凤凰人,硕士生,主要从事禽病与病原分子生物研究,E-mail:cuilanwu@163.com
  • 基金资助:

    国家自然科学基金项目(31360611;31160516);广西自然科学基金项目(2014GXNSFDA118011;2013GXNSFCA019010)

Isolation and Analysis of Structural Protein Genes Variation and Serotype of a Recombinant Strain of Chicken Infectious Bronchitis Virus

WU Cui-lan,HE Yi-ning,LI He-ming,SUN Xin-kuan,TAN Yu-cai,WEI Tian-chao,MO Mei-lan*,WEI Ping*   

  1. (College of Animal Science and Technology,Guangxi University,Nanning 530005,China)
  • Received:2014-09-10 Online:2015-05-23 Published:2015-05-23

摘要:

为进一步了解鸡传染性支气管炎病毒(IBV)流行株的遗传变异情况,对2013年从免疫失败的广西某鸡场中分离到的一株IBV(GX-NN130048)进行S1、EMN基因的扩增、测序、相似性比较、系统进化树绘制和重组分析以及血清型鉴定。基因序列分析结果表明,分离株GX-NN130048的S蛋白裂解位点序列为RRSRR,S1、EMN基因与参考株核苷酸相似性分别在60.8%~90.1%、81.0%~94.2%、86.1%~93.8%和86.0%~93.3%;进化树分析显示,分离株GX-NN130048的S1基因与疫苗毒株4/91同属一群,而EMN基因则与毒株LX4属于同一群;重组分析显示,分离株GX-NN130048的S1基因来源于疫苗毒株4/91和GX-HC1006野毒株(LX4型)的重组;血清型分析表明,该分离株的血清型不同于疫苗株H120和4/91。研究表明分离株GX-NN130048是一株重组毒株,其结构基因和血清型均发生变异。本研究再次提供证据说明,疫苗株4/91越来越多参与到IBV流行株通过基因重组导致的遗传变异事件当中,并可能由此导致新血清型的出现。这可能是现场免疫失败的一个主要原因。有必要继续加强对IBV的监测和新疫苗的研发。

Abstract:

In order to further understand the genetic variation of circulating infectious bronchitis virus(IBV),S1,EM and N genes of an IBV strain(GX-NN130048),which was isolated from chickens with immunoprophylaxis defeat in Guangxi,were amplified by reverse transcriptase polymerase chain reaction(RT-PCR),and then cloned,sequenced,and similarity comparison,phylogenetic tree,recombination and serotype analysis were conducted.The results showed that the gene sequence of cleavage site within S protein of GX-NN130048 was RRSRR.The nucleotide similarities of S1,EM and N genes with those of other reference strains were 60.8%-90.1%,81.0%-94.2%,86.1%-93.8% and 86.0%-93.3%,respectively.According to the phylogenetic tree analysis,it was clustered into 4/91-type based on S1 gene,while its EM and N genes belonged to LX4-type.Recombination event was detected in S1 gene,which was a recombinant between vaccine strain 4/91 and isolate GX-HC1006(LX4-type).The serotype analysis revealed that the serotype of GX-NN130048 was different from vaccine strains H120 and 4/91.The results indicated that isolate GX-NN130048 was a recombinant strain and variation existed both in structural genes and serotype.Our results also provide the evidence that the vaccine strain 4/91 has been increasingly involved in genetic variation of IBV epidemic strains,which may lead to the emergence of new serotype,and that may be one of the main causes of vaccine break in the field.Therefore,it is necessary to strengthen the monitoring of IBV field isolates and development of new vaccines.

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