| 应用TaqMan-BHQ探针实时荧光RT-PCR法定量检测马秋波病毒核酸 |
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| 引用本文： | 燕清丽,张晓龙,魏莲,姚李四. 应用TaqMan-BHQ探针实时荧光RT-PCR法定量检测马秋波病毒核酸[J]. 中国国境卫生检疫杂志, 2010, 0(3): 149-152 |
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| 作者姓名： | 燕清丽 张晓龙 魏莲 姚李四 |
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| 作者单位： | 中国检验检疫科学研究院;军事医学科学院; |
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| 基金项目： | 中国检验检疫科学研究院院所长基金(2008JK009) |
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| 摘 要： | 〔目的〕建立一种适应口岸马秋波病毒实时荧光定量RT-PCR快速的检测方法。〔方法〕用专业软件设计引物和TaqMan-BHQ探针,以人工合成马秋波病毒S基因的片段作为模板,进行实时荧光定量RT-PCR研究。〔结果〕模板的Ct值与模板稀释浓度的对数存在良好的线性关系,标准曲线为Y=-3.281X+50.975,R2=0.999361,PCR扩增效率为101.0%,其最低检出限为28 copies/μl。〔结论〕应用TaqMan-BHQ1探针的实时荧光RT-PCR检测马秋波病毒核酸,具有耗时短、灵敏度高等特点。
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| 关 键 词： | TaqMan-BHQ探针 实时荧光RT-PCR 马秋波病毒 定量检测 应用 |
Application of TaqMan-BHQ Probe on Quantitative Detection for Machupo Virus by Real-time Fluorescence RT-PCR |
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| Yan Qingli,Zhang Xiaolong,Wei Lian,Yao Lisi. Application of TaqMan-BHQ Probe on Quantitative Detection for Machupo Virus by Real-time Fluorescence RT-PCR[J]. Chinese Journal of Frontier Health and Quarantine, 2010, 0(3): 149-152 |
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| Authors: | Yan Qingli Zhang Xiaolong Wei Lian Yao Lisi |
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| Affiliation: | Yan Qingli1,Zhang Xiaolong1,Wei Lian1,Yao Lisi1,2(1.Chinese Academy of Inspection and Quarantine,Beijing 100123,China,2.The Academy of Military Medical Science,Beijing 100071,China.) |
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| Abstract: | Objective To establish a rapid and accurate method of real-time fluorescence RT-PCR to quantify the S gene of Machupo virus.Methods The twin primers and the TaqMan-BHQ probe were designed and synthesized based on the S gene segment of Machpo virus.The real-time RT-PCR was developed for detecting machupo virus.Results The Ct value of templates had a linear relationship with the log starting quantity.Sensitivity assay showed that the detection limit of the assay was 28 copies/μl and the standard curve Y=-3.281X＋50.975 had a good reproducibility.Conclusions Quantitative real-time fluorescence for Machupo virus detection was established and characterized by rapidity and sensitivity. |
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| Keywords: | TaqMan-BHQ probe Real-time fluorescence RT-PCR Machupo virus Quantitative detection Application |
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