| A型H1N1及H3N2亚型流感病毒实时荧光RT-PCR检测方法的建立 |
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| 引用本文： | 杨素,陈博文,沙才华,廖明,秦爱建,徐海聂,廖秀云,罗宝正,薄清如,陈伟生.A型H1N1及H3N2亚型流感病毒实时荧光RT-PCR检测方法的建立[J].中国兽医科学,2010,40(3). |
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| 作者姓名： | 杨素 陈博文 沙才华 廖明 秦爱建 徐海聂 廖秀云 罗宝正 薄清如 陈伟生 |
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| 作者单位： | 杨素(扬州大学,兽医学院,江苏,扬州,225009;珠海出入境检验检疫局,广东,珠海,519015);陈伟生,陈博文,沙才华,徐海聂,廖秀云,罗宝正,薄清如(珠海出入境检验检疫局,广东,珠海,519015);廖明(华南农业大学,兽医学院,广东,广州,510642);秦爱建(扬州大学,兽医学院,江苏,扬州,225009) |
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| 基金项目： | 广东省农业攻关项目，广东省农产品安全生产关键技术研究与示范重点专项，珠海市科技项目，珠海出入境检验检疫局科技项目 |
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| 摘 要： | 根据A型H1N1亚型流感病毒2009年北美变异株和H3N2亚型流行株的基因序列,设计了3套特异性引物、TaqMan探针,分别用不同的荧光基团标记,以构建的A型流感病毒及H1N1和H3N2亚型流感病毒基因重组质粒作为阳性对照,经反应条件优化,特异性、敏感性和重复性试验,筛选出3套不同的反应体系及同一个反应参数,建立了可同时检测A型流感病毒、H1N1和H3N2亚型流感病毒的实时荧光RT-PCR方法。结果表明,该方法可特异地检测H1N1(人源、猪源)、H3N2(猪源),禽流感病毒H5、H9等A型流感病毒,且与新城疫病毒、减蛋综合征病毒等其他病毒不发生交叉反应,具有特异、敏感、重复性好、快速、费用低廉等优点,试验耗时仅3h。表明,建立的方法,一次试验即可完成A型流感病毒的初筛和H1N1、H3N2亚型流感病毒的初步确认定型。
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| 关 键 词： | A型流感病毒 H1N1亚型 H3N2亚型 实时荧光RT-PCR |
Development of real-time RT-PCR for simultaneous detection of influenza A virus and subtypes of H1N1 and H3N2 |
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| YANG Su,CHEN Bo-wen,SHA Cai-hua,LIAO Ming,QIN Ai-jian,XU Hai-nie,LIAO Xiu-yun,LUO Bao-zheng,BO Qing-ru,CHEN Wei-sheng.Development of real-time RT-PCR for simultaneous detection of influenza A virus and subtypes of H1N1 and H3N2[J].Veterinary Science in China,2010,40(3). |
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| Authors: | YANG Su CHEN Bo-wen SHA Cai-hua LIAO Ming QIN Ai-jian XU Hai-nie LIAO Xiu-yun LUO Bao-zheng BO Qing-ru CHEN Wei-sheng |
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| Institution: | YANG Su1,2,CHEN Bo-wen2,SHA Cai-hua2,LIAO Ming3,QIN Ai-jian1,XU Hai-nie2,LIAO Xiu-yun2,LUO Bao-zheng2,BO Qing-ru2,CHEN Wei-sheng2(1.College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China,2.Zhuhai Entry-Exit Inspection & Quarantine Bureau,Zhuhai 519015,3.College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China) |
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| Abstract: | To develop a real-time RT-PCR for preliminary detection and differentiation of influenza A virus and subtypes H1N1 and H3N2 at one test,based on gene nucleotide sequences of Influenza A Virus/H1N1/2009 strain and Influenza A Virus/H3N2 strain in GenBank,three sets of specific primers and TaqMan probes were designed and labelled with different fluorescence reporters respectively,their recombinant plasmids were constructed to be used as positive control,and three different reaction mixtures and same RT-PCR co... |
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| Keywords: | influenza A virus H1N1 subtype H3N2 subtype real-time RT-PCR |
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