›› 2011, Vol. 38 ›› Issue (10): 94-98.

• 生物技术 • 上一篇    下一篇

奶牛传染性鼻气管炎病毒gG基因PCR检测方法的建立

王冰1,2,3, 张敏敏1,3, 邹新峰1,3, 郭爱珍1,3   

  1. 1. 华中农业大学农业微生物学国家重点实验室,湖北武汉 430070;2. 华中农业大学动物科技学院,湖北武汉 430070;3. 华中农业大学动物医学院,湖北武汉 430070
  • 收稿日期:1900-01-01 修回日期:2011-03-30 出版日期:2011-10-20 发布日期:2011-10-20
  • 通讯作者: 郭爱珍

Establishment of PCR Method to Detect gG Gene of Infectious Bovine Rhinotracheitis Virus

WANG Bing1,2,3, ZHANG Min-min1,3, ZOU Xin-feng1,3, GUO Ai-zhen1,3   

  1. 1. The State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070, China;2. College of Animal Science, Huazhong Agricultural University,Wuhan 430070, China;3. College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070, China
  • Received:1900-01-01 Revised:2011-03-30 Online:2011-10-20 Published:2011-10-20

摘要: 本试验旨在建立一种PCR技术,既能快速检测牛传染性鼻气管炎病毒,又能区分同属病毒牛疱疹病毒5型和伪狂犬病病毒。根据基因库中牛传染性鼻气管炎病毒gG基因的特异性引物,建立PCR方法,对牛传染性鼻气管炎病毒参考毒株和阳性样本进行扩增,结果均能扩增出一条463 bp的特异性条带;对同属的牛疱疹病毒5型进行扩增,获得651 bp和431 bp两条带;对同属伪狂犬病病毒进行扩增,获得493 bp的条带;而非相关病毒(如猪呼吸与繁殖综合征病毒等),不能扩增出条带。对牛传染性鼻气管炎病毒的检测灵敏度为2×10-3 PFU/mL。鉴于该方法具有良好的灵敏度和特异性,将在牛疱疹病毒感染诊断和标记疫苗免疫后的鉴别诊断方面具有良好的应用前景。

关键词: 牛传染性鼻气管炎病毒; PCR; 牛疱疹病毒; gG基因

Abstract: The specific primers were designed according to the published sequence of the infectious bovine rhinotracheitis virus (IBRV)gG gene. The method could amplify a specific IBRV fragment of 463 bp for IBRV reference strain and the IBRV positive samples. For bovine herpesvirus 5,it amplified two bands of 651 bp and 431 bp;for pseudorabies,the PCR product is a fragment of 493 bp. However,this PCR did not yield any product for non-related viruses such as porcine reproductive and respiratory syndrome virus. The sensitivity for IBRV was 2×10-3 PFU/mL. Since this PCR has good sensitivity and specificity,it would have a wide application in diagnosis of bovine herpesvirus infection and differential detection among the members of herpesviruses and between natural infection and vaccination of gG deleted marker vaccine. A PCR method was established successfully to detect infectious bovine rhinotracheitis virus (IBRV) and differentiate bovine herpesviruses.

Key words: infectious bovine rhinotracheitis virus (IBRV); PCR; bovine herpesvirus; gG gene

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