| 甲型H1N1流感病毒核酸荧光定量RT-PCR检测技术 |
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| 引用本文： | 尹惠琼,刘松婷,史利军,杨姝,章金刚.甲型H1N1流感病毒核酸荧光定量RT-PCR检测技术[J].生物技术通讯,2010,21(2):244-247. |
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| 作者姓名： | 尹惠琼 刘松婷 史利军 杨姝 章金刚 |
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| 作者单位： | 军事医学科学院,野战输血研究所,北京,100850 |
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| 摘 要： | 目的:建立基于TaqMan荧光探针技术的甲型H1N1流感病毒实时定量RT-PCR方法。方法:分析H1N1流感病毒基因特性,根据新发甲型H1N1流感病毒突变基因片段设计检测引物和TaqMan荧光探针,建立荧光定量RT-PCR检测体系,体外转录制备RNA标准品,进行特异性、敏感性、重复性及盲样检测评价实验。结果:可特异性有效检测新发甲型H1N1流感病毒核酸,与H1～H16流感病毒基因无交叉反应;对RNA标准品的检测敏感性达103拷贝/μL;重复性实验中,阳性标准品Ct值变异系数(CV)10%,阴性标准品检测结果均呈阴性;12份盲样检测结果特异性好。结论:该荧光定量RT-PCR方法可作为甲型H1N1流感防控的病原快速诊断技术。
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| 关 键 词： | 甲型H1N1流感病毒 大流行 荧光定量RT—PCR 诊断 |
Detection and Quantification of Novel H1N1 Influenza A Virus with TaqMan Probe Based Real-Time RT-PCR |
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| YIN Hui-Qiong,LIU Song-Ting,SHI Li-Jun,YANG Shu,ZHANG Jin-Gang.Detection and Quantification of Novel H1N1 Influenza A Virus with TaqMan Probe Based Real-Time RT-PCR[J].Letters in Biotechnology,2010,21(2):244-247. |
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| Authors: | YIN Hui-Qiong LIU Song-Ting SHI Li-Jun YANG Shu ZHANG Jin-Gang |
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| Institution: | YIN Hui-Qiong,LIU Song-Ting,SHI Li-Jun,YANG Shu,ZHANG Jin-Gang Viral Safety Laboratory of National Center of Biomedical Analysis,Institute of Transfusion Medicine,Academy of Military Medical Sciences,Beijing 100850,China |
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| Abstract: | Objective:To develop a novel quantitative real-time RT-PCR based on TaqMan fluorescent probe detection for new H1N1 influenza A virus which produced pandemic in 2009.Methods:The gene sequences of hemagglutinin(HA) of influenza A virus were aligned by using bioinformatics software,and a set of primers and a TaqMan fluorescent probe were designed,and analyzed through BLAST online.Standard RNA was produced through T7 RNA polymerase in vitro.Specificity,sensitivity and repeatability of the assay were assessed.B... |
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| Keywords: | influenza A(H1N1) virus pandemic real-time RT-PCR diagnose |
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