鸡传染性支气管炎病毒变异株ZZ2004 RT-PCR检测方法的建立及应用  

Establishment and application of RT-PCR technique for detection of the ZZ2004 serotype of infectious bronchitis virus

作　　者：姚四新[1,2] 王宪文[1] 赵淑秋[1] 刘兴友[1,2] 

机构地区：[1]河南科技学院动物科学学院,河南新乡453003 [2]动物疫病和残留物防控河南省高校工程技术研究中心,河南新乡453003

出　　处：《中国兽医杂志》2013年第4期15-17,共3页Chinese Journal of Veterinary Medicine

基　　金：国家自然科学基金(30170707);2012年河南教育厅科学技术研究重点项目(12A230004);河南科技学院院级重点基金项目(2011013)

摘　　要：根据GenBank已发表的鸡传染性支气管炎病毒(Infectious bronchitis virus,IBV)ZZ2004 3a、3b及E基因序列,设计1对引物,扩增540bp特异性核酸片段,建立了检测IBV ZZ2004的RT-PCR方法。特异性试验结果表明,IBV ZZ2004能扩增出540bp的核酸片段,而IBV M41、H120、H52毒株以及新城疫病毒(NDV)、禽流感病毒(AIV)、传染性腔上囊病病毒(IBDV)均无特异性条带出现。敏感性试验结果表明,该方法的最低检出量为1.525pg的模板。上述结果表明,本试验所建立的RT-PCR方法敏感性高、特异性强。利用建立的RT-PCR方法对从河南省分离的6株疑似鸡IBV ZZ2004进行检测,结果5株为阳性。该方法的建立为IBV ZZ2004的诊断及流行病学调查提供了可靠的方法。According to the sequences of the ZZ2004 serotype of IBV 3a, 3b and E gene published in GenBank, a pair of primers was designed and synthesized to specifically amplify the 540 bp nucleotide fragment, the RT -PCR technique for detection of the ZZ2004 serotype of IBV was established. The 540 bp specific strip was found only in the PCR amplification of the ZZ2004 serotype of IBV in the specificity assay, the same strip didn＇t appear in other viruses and strains of IBV. The sensitivity result＇indicated that as little as 1.525 pg in the sample were detected in PCR. Five of the six uncertain isolates detected by the established RT -PCR technique were positive. The above results showed that the technique provided a more sensitive, specific and rapid method for diagnosis and epidemio- logical survey of the ZZ2004 serotype of IBV.

关 键 词：传染性支气管炎病毒 RT-PCR 鸡 

分 类 号：S852.659.6[农业科学—基础兽医学]