期刊文献+

上海地区鸡传染性支气管炎病毒S2基因的检测及分析 被引量:2

Molecular Epidemics of Infectious Bronchitis Virus in Shanghai District and Analysis of Its S2 Fragment
在线阅读 下载PDF
导出
摘要 参考GenBank发表的传染性支气管炎病毒(infectious bronchitis virus,IBV)S基因序列,设计合成了1对引物,模拟PCR表明,该引物能扩增现已发表的所有IBV的S2基因部分核酸序列,扩增片段493bp。建立的RT—PCR检洲体系检测灵敏度达10~50ELD50,检出限量为0.1ng。采集上海地区12个鸡场疑似IBV禽样品34份进行检洲,阳性为32份,序列分析表明均为肾变型IBV S2基因片段,S2基因同源性较高(≥99%),与其他地区肾变型IBV也有较高同源性,而与腺胃型和呼吸型相比较,序列差异较大。进化分析表明,尽管IBV S2片段变异率明显低于S1基因,但二者表现相同的进化趋势,S2基因亦可反映出IBV的分子变异及毒株间的亲缘关系。 A pair of specific primer was designed and synthesized in the conserved region of S2 gene according to the infectious bronchitis virus(IBV) S sequences at GenBank. A 493 bp amplicon was abtained from simulating PCR using software SPCR3.0. The PCR system was specific with IBV while no reaction with other avian virus tested in the study. 10-50 ELD50 of the viron was detected. By using this method to detect the clinical samples,which were collected from 12 chicken factories,32 out of 34 suspicious samples were positive. Further analysis showed all the IBV-positive belonged to the nephropathogenic viruses. The S2 genes of the epidemic viruses share high identities with each other,while has much more difference with the proventriculopathogenic IBV and other viruses causing respiratory symptom in China. Phylogenic analysis showed though S2 fragments have less divergence among isolates than S1 genes,it shows the same phylogenic tendency with S gene among the viruses.
出处 《中国兽医学报》 CAS CSCD 北大核心 2006年第2期126-128,132,共4页 Chinese Journal of Veterinary Science
基金 上海市农委重点攻关项目
关键词 鸡传染性支气管炎 S2基因 进化分析 avian infectious bronchitis virus S2 gene phylogenic analysis
  • 相关文献

参考文献10

  • 1陆承平.兽医微生物学[M].第3版.北京:中国农业出版社,2000.548-554.
  • 2周继勇,张德勇,丁红梅,程丽琴,陈吉刚.不同组织嗜性传染性支气管炎病毒核衣壳基因的遗传变异性分析[J].病毒学报,2003,19(1):59-63. 被引量:5
  • 3周继勇.传染性腺胃炎病毒ZJ_(971)株的一些生物学特性[J].畜牧兽医学报,2000,31(3):229-234. 被引量:27
  • 4Chomczynaki P Sacchin.Single step method of RNA isolation by acid guanidinium thiocyanate-hpenol-chloroform extration[J].Anal Biochem,1987,162:156-159.
  • 5萨姆布鲁克J 弗里奇EF 曼尼阿蒂斯T著.金冬雁 黎孟枫 译.分子克隆实验指南[M].第2版[M].北京:科学出版社,1992.456-494.
  • 6Jack Wood M W,Hilt D A,Callison S A,et al.Detection of infectious bronchitis virus by real-time reverse transcprtase-polymerase chain reaction and ifentification of a quasispecies in the Beaudette stain[J].Avian Dis,2003,9:47 137.
  • 7Liu H J,Lee L H,Shih W L,et al.Detection of infectious bronchitis virus by multiplex polymerase chain reaction and sequence analysis[J].J Virol Methods,2003,109 (1):31-37.
  • 8Pang Y,Wang H,Girshick T,et al.Development and application of a multiplex polymerase chain reaction for avian respiratory agents[J].Avian Dis,2002,46(3):691-699.
  • 9Thomas M Gallagher,Michae J B.Coronavirus spike protein in viral entry and pathogenesis[J].Virology,2001,279:371-374.
  • 10Koch G,Hartog L,Kant A,et al.Antigenic domains on the peplomer protein of avian infectious fronchitis virus:correlation with biological functions[J].J Gen Viro,1990,71:1 929-1 935.

二级参考文献11

共引文献51

同被引文献21

  • 1[1]Emiliana F,Emanuela D A,Livia D T,et al.Rapid diagnosis of avian infectious bronchitis virus by polymerase chain reaction[J].J Virol Meth,1997,64 (2):125-130.
  • 2[3]Cavanagh D.A nomenclature for avian coronavirus isolates and the question of species status[J].Aian Pathol,2001,30 (2):109-115.
  • 3[4]Wit J J de,Koch G,Kant A,et al.Detection by immunofluorescent.Assay of serotype specific and group specific antigens of infectious bronchitis virus in tracheas of broilers with respiratory problems[J].Avian Pathol,1995,24 (5):465-474.
  • 4[5]Stephensen C B,Casebolt D B,Gangopadhyay N N.Phylogenetic analysis of a highly conserved region of the polymerase gene from 11 coronaviruses and development of a consensus polymerase chain reaction assay[J].Virus Res,1999,60 (2):181-189.
  • 5[6]Chomczynaki P S.Single step method of RNA iso-lation by acid guanidinium thiocyanate-phenol-chloroform extration[J].Analyt Biochem,1987,162 (1):156-159.
  • 6[7]Carr W H,Sweeney J N,Nunan L,et al.The use of an infectious hypodermal and hematopoietic necrosis virus gene probe serodiag nostic field kit for the screening of candidate specific pathogen-free Penaeus vannamei broodstock[J].Aquaculture,1996,147 (1-2):1-8.
  • 7[10]Poulos B T,Mari J,Bonami J R,et al.Use of non-radioactive labeled DNA probes for the detection of a baculovirus from penaeus monodon by in situ hybridization on fixed tissue[J].J Virol Methods,1994,49 (2):187-194.
  • 8[11]Nunan L M,Lightner D L.Development of a nonradioactive DNA probe by PCR for detection of white spot syndrome virus (WSSV)[J].J Virol Methods,1997,63 (1-2):193-201.
  • 9Allander T,Emerson SU,Engle RE,et al.A virus discovery method incorporating DNase treatment and its application to the identification of two bovine parvovirus species.Proceedings of the National Academy of Sciences of the United States of America,2001,98(20):11609-11614.
  • 10Van der Hoek L,Pyrc K,Jebbink MF,et al.Identification of a new human coronavirus.Nature medicine,2004,10(4):368-373.

引证文献2

二级引证文献5

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部