| H9亚型禽流感病毒和鸭坦布苏病毒二重PCR检测方法的建立 |
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| 引用本文： | 赵虹,谢芝勋,谢丽基,刘加波,邓显文,谢志勤,庞耀珊,范晴,罗思思.H9亚型禽流感病毒和鸭坦布苏病毒二重PCR检测方法的建立[J].中国畜牧兽医,2014,41(10):43-47. |
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| 作者姓名： | 赵虹 谢芝勋 谢丽基 刘加波 邓显文 谢志勤 庞耀珊 范晴 罗思思 |
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| 作者单位： | 1. 广西大学动物科学技术学院, 广西南宁 530004;2. 广西壮族自治区兽医研究所, 广西畜禽疫苗新技术重点实验室, 广西南宁 530001 |
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| 基金项目： | 广西特聘专家专项经费(2011B020);广西科技攻关重大专项(1222003-2-4);广西科技攻关项目(桂科攻10100014-5);广西重点实验室建设项目(12-071-28-A-3);广西重点实验室建设项目(12-071-28-A-4) |
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| 摘 要： | 本试验旨在建立一种能同时快速检测H9亚型禽流感病毒(avian influenza virus,AIV)和鸭坦布苏病毒(duck Tembusu virus,DTMUV)的二重PCR检测方法。根据GenBank中H9亚型 AIV的HA基因和DTMUV的NS2基因序列,分别设计了1对针对H9亚型AIV和DTMUV保守基因序列的引物。利用这2对引物对混有H9亚型AIV和DTMUV的cDNA模板进行二重PCR扩增,得到了2个大小与试验设计相符的特异性扩增条带。利用本试验建立的二重PCR对其他鸭病病原体进行PCR扩增,结果均为阴性。利用这2对引物对H9亚型AIV和DTMUV进行敏感性检测,结果显示最低检测极限分别为6.3和 6.6 pg。对临床235份样品检测的结果表明该二重PCR检测方法具有快速、敏感、特异等优点,适用于临床检测的应用。
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| 关 键 词： | H9亚型禽流感病毒 鸭坦布苏病毒 二重PCR |
| 收稿时间： | 2014-05-08 |
Establishment of Duplex PCR Assay for Detection of H9 Subtype Avian Influenza Virus and Duck Tembusu Virus |
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| ZHAO Hong,XIE Zhi-xun,XIE Li-ji,LIU Jia-bo,DENG Xian-wen,XIE Zhi-qin,PANG Yao-shan,FAN Qing,LUO Si-si.Establishment of Duplex PCR Assay for Detection of H9 Subtype Avian Influenza Virus and Duck Tembusu Virus[J].China Animal Husbandry & Veterinary Medicine,2014,41(10):43-47. |
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| Authors: | ZHAO Hong XIE Zhi-xun XIE Li-ji LIU Jia-bo DENG Xian-wen XIE Zhi-qin PANG Yao-shan FAN Qing LUO Si-si |
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| Institution: | 1. College of Animal Science and Technology, Guangxi University, Nanning 530004, China;2. Guangxi Key Laboratory of Animal Vaccines and New Technology, Guangxi Veterinary Research Institute, Nanning 530001, China |
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| Abstract: | The assay was aimed to establish a duplex PCR assay to detect H9 subtype avian influenza virus (AIV) and duck Tembusu virus (DTMUV) rapidly and simultaneously. According to the sequences of H9 AIV HA gene and DTMUV NS2 gene in GenBank,two sets of specific primers were designed. It was shown that all samples containing H9 subtype AIV and DTMUV could be amplified into two specific bands,but no specific bands were amplified from other duck disease virus. The sensitive determination result indicated that as little as 6.3 pg of H9 subtype AIV and 6.6 pg of DTMUV could be detected by this duplex PCR.Finally,based on 235 samples of clinical test indicated that this duplex PCR assay with the advantages of quick,sensitive and specific,could be useful for the clinical detection. |
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| Keywords: | H9 subtype avian influenza virus duck Tembusu virus duplex PCR |
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