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鸡传染性支气管炎病毒793/B RT-PCR检测方法的建立及应用

Establishment and application of RT-PCR technigue for detection of the 793/B serotype of infectious bronchitis virus

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【作者】 于申业刁有祥杨杰华高晓伟刘霞刘悦竹陈庆普

【Author】 YU Shen-ye1,4,DIAO You-xiang1,YANG Jie-hua1,GAO Xiao-wei2,LIU Xia1,LIU Yue-zhu3,CHEN Qing-pu3 (1.College of Animal Science and Technology,Shandong Agricultural University,Taian, Shandong 271018,China;2.Agricultural College of Jilin University,Changchun 130062,China;3.Mingfa Animal Pharmacal Co.Ltd.of Shandong Province,Ji′nan 250022,China;4.Zoonoses Institute,Jilin University,Changchun 130062,China)

【机构】 山东农业大学动物科技学院吉林大学农学部山东明发兽药股份有限公司山东明发兽药股份有限公司 山东泰安271018吉林大学人兽共患病研究所吉林长春130062山东泰安271018山东济南250036

【摘要】 根据GenBank已发表的鸡传染性支气管炎病毒(Infectious bronchitis virus,IBV)793/BS1基因序列,设计1对引物,扩增891bp特异性核酸片段,建立了检测IBV793/B的RT-PCR方法。特异性试验结果表明,IBV793/B能扩增出891bp的核酸片段,而IBVM41、H120、H52毒株以及新城疫病毒(NDV)、禽流感病毒(AIV)、传染性法氏囊病病毒(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10pg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方法对从山东省分离的8株疑似鸡IBV793/B进行检测,结果7株为阳性。该方法的建立为IBV793/B的诊断及流行病学调查提供了可靠的方法。

【Abstract】 According to the sequences of the 793/B serotype of IBV S1 gene published in GenBank,a pair of primers was designed and synthesized to specifically amplify the 891 bp nucleotide fragment,the RT-PCR technique for detection of the 793/B serotype of IBV was established.The 891 bp special strip was found only in the PCR amplification of the 793/B serotype of IBV in the specificity assay,there were no the same strip appeared in the other viruses and strains of IBV.The sensitivity result indicated that as little as 10 pg in sample were detected in PCR.Seven of the eight 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 epidemiological survey of the 793/B serotype of IBV.

【基金】 济南市科技攻关项目(41054)
  • 【文献出处】 中国兽医学报 ,Chinese Journal of Veterinary Science , 编辑部邮箱 ,2007年04期
  • 【分类号】S854.43
  • 【被引频次】7
  • 【下载频次】175
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