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禽流感病毒H5,H7,H9亚型多重RT-PCR检测方法的建立

Construction of multiplex RT-PCR for detection of H5,H7 and H9 subtypes of avian influenza virus

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【作者】 胡建和赵坤杭柏林荆汝顶王丽荣王三虎刘兴友

【Author】 HU Jian-he1,ZHAO Kun1,HANG Bo-lin1,JING Ru-ding2,WANG Li-rong1,WANG San-hu1,LIU Xing-you1(1.School of Animal Science,Henan Institute of Science and Technology,Xinxiang 453003,China;2.Animal Husbandry and Veterinary Bureau of Xinxiang,Xinxiang 453003,China)

【机构】 河南科技学院动物科学学院新乡市畜牧兽医工作站

【摘要】 依据GenBank中亚洲流行株禽流感病毒(AIV)H5,H7,H9亚型血凝素基因及M基因序列保守区域设计特异性引物,利用多重反转录聚合酶链式反应技术建立了AIV及H5,H7,H9亚型的检测方法.该方法能一次性检测出AIV保守区域M基因的同时,直接区分H5,H7,H9亚型.对新城疫病毒、传染性法氏囊病毒、传染性支气管炎病毒、传染性喉气管炎病毒等特异性检测均为阴性.样品经100倍稀释后仍能检出.

【Abstract】 A multiplex RT-PCR was constructed to detect H5,H7,H9 subtypes of avian influenza virus.Four sets of specific oligonucleotide primers were designed for detection and differentiation of H5,H7 and H9 subtypes of avian influenza virus,which was based on sequences of hemagglutinin gene and matrix gene of avian influenza virus from GenBank.This method could detect matrix gene of AIV,and simultaneously identify H5,H7 and H9 subtypes.Detections of newcastle disease virus,infectious bursal disease virus,infectious bronchitis virus,infectious laryngotracheitis virus were all negative.It could detect RNA that was diluted 100 times.So it is an excellent method to diagnose and differentiate H5,H7 and H9 subtypes of AIV.

【基金】 河南省高校杰出科研人才创新工程项目(2006KYC020)
  • 【文献出处】 河南农业大学学报 ,Journal of Henan Agricultural University , 编辑部邮箱 ,2009年02期
  • 【分类号】S854.43
  • 【被引频次】1
  • 【下载频次】223
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