应用TaqMan-BHQ探针实时荧光RT-PCR法定量检测马秋波病毒核酸

    应用TaqMan-BHQ探针实时荧光RT-PCR法定量检测马秋波病毒核酸

    燕清丽1 张晓龙1 魏 莲1 姚李四1，2

    （1.中国检验检疫科学研究院，北京 100123；2.军事医学科学院，北京100071）

    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。

    关键词 TaqMan-BHQ探针;实时荧光RT-PCR;马秋波病毒；定量检测；应用

    〔中图分类号〕　R512.8 R184.35　〔文献标识码〕　B

    Application of TaqMan-BHQ Probe onQuantitative Detection for Machupo Virus by Real-time FluorescenceRT-PCR  Yan Qingli1, Zhang Xiaolong1,Wei Lian1,Yao Lisi1,2(1.Chinese Academy of Inspection and Quarantine, Beijing100123,China; 2.The Academy of Military Medical Science, Beijing100071,China.)

    [Abstract]   Objective  To establish a rapid and accurate method of real-time fluorescenceRT-PCR to quantify the S gene of Machupo virus. Methods  Thetwin primers and the TaqMan-BHQ probe were designed and synthesizedbased on the S gene segment of Machpo virus. The real-time RT-PCRwas developed for detecting machupo virus. Results   The Ct value of templates had a linearrelationship with the log starting quantity. Sensitivity assayshowed that the detection limit of the assay was 28 copies/μl andthe standard curve Y=-3.281X+50.975 had a goodreproducibility.  Conclusions   Quantitativereal-time fluorescence for Machupo virus detection was establishedand characterized by rapidity and sensitivity.

    [Key words]   TaqMan-BHQ probe;Real-time fluorescence RT-PCR; Machupo virus; Quantitativedetection; Application