[1]尹 航,杨焕良,陈 艳,等.甲型H1N1流感病毒实时荧光定量PCR检测方法的建立[J].中国预防兽医学报,2013,(01):36.
 YIN Hang,YANG Huan-liang,CHEN Yan,et al.Establishment of one-step TaqMan real time RT-PCR for Pandemic/2009 influenza viruses in swine[J].Chinese journal of preventive veterinary medicine,2013,(01):36.
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甲型H1N1流感病毒实时荧光定量PCR检测方法的建立
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2013年01期
页码:
36
栏目:
诊断和检测技术
出版日期:
2013-01-15

文章信息/Info

Title:
Establishment of one-step TaqMan real time RT-PCR for Pandemic/2009 influenza viruses in swine
作者:
尹 航12杨焕良1陈 艳1孟沙沙1刘丽萍1辛晓光1陈化兰1乔传玲1*
(1. 中国农业科学院哈尔滨兽医研究所 农业部动物流感重点开放实验室/兽医生物国家重点开放实验室,黑龙江 哈尔滨 150001;2. 东北农业大学,黑龙江 哈尔滨 150030)
Author(s):
YIN Hang12 YANG Huan-liang1 CHEN Yan1 MENG Sha-sha1 LIU Li-ping1 XIN Xiao-guang1 CHEN Hua-lan1 QIAO Chuan-ling1*
(1. Animal Influenza Key Laboratory of the Ministry of Agricultural, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China; 2. Northeast Agricultural University, Harbin 150030, China)
关键词:
猪流感病毒H1N1亚型TaqMan MGB探针实时荧光定量PCR
Keywords:
swine influenza virus H1N1 subtype TaqMan MGB probe one-step real time PCR
摘要:
为建立快速检测甲型H1N1流感病毒[Influenza A (H1N1) pdm09]的TaqMan荧光定量PCR方法,本研究根据甲型H1N1流感病毒血凝素(HA)基因保守序列,设计并合成了特异性引物和TaqMan MGB探针,采用实时荧光定量PCR技术检测甲型H1N1流感病毒。使用含有选定检测序列的重组质粒pMD-HA标准品绘制标准曲线,其相关系数为0.999,敏感性可达50 TCID50。本研究中,甲型H1N1流感病毒TaqMan荧光定量RT-PCR方法与经典H1N1、类禽H1N1、类人H1N1、H1N2、H3N2、H5N1和H9N2流感病毒无交叉反应。结果表明,该方法可以在人类和动物间有效地检测甲型H1N1流感病毒。
Abstract:
To establish detect method for the Pandemic/2009 influenza virus infection in swine, A TaqMan real-time RT-PCR was developed based on primers and TaqMan probe derived from pH1N1/09 HA gene. A serial dilution of recombinant plasmid of pMD-H1-HA was prepared and used to generate standard curves. The results showed that the sensitivity of the real-time RT-PCR was 50 TCID50, with a correlation coefficient of 0.999 and no cross-reaction with classical swine H1, Eurasian avian-like H1, human-like H1, or H1N2, H3N2, H5N1, H9N2 influenza viruses. The results demonstrate that this assay is efficient for the diagnosis of pH1N1/09 virus in both human and animal.

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更新日期/Last Update: 2013-01-20