裂谷热病毒荧光定量PCR检测方法的建立

    杨志红1，2，李长田1，杨宇2，白琳2，魏莲2，王静2

    1.吉林农业大学中药材学院，吉林 长春 130012；2.中国检验检疫科学研究院

    摘要：目的建立一种快速、敏感、特异的实时荧光定量PCR方法，用于裂谷热病毒的检测。方法　通过序列对比在裂谷热病毒L基因保守区设计引物及Taqman探针，建立实时荧光定量PCR反应体系。结果经优化的荧光定量PCR方法有较好的灵敏度和特异性，对阳性对照质粒标准品的灵敏度可达37拷贝/μl，通过检测同为蚊媒传播的日本脑炎病毒、黄热病毒、登革病毒、基孔肯亚病毒无交叉反应。结论本方法的建立在国境口岸传染病的防控方面有较好的应用前景。

    关键词：荧光定量PCR；检测；裂谷热

    中图分类号：R535　文献标识码：B

    Real-time PCR method for rapid detection ofRift Valley Fever virus

    YANG Zhi-hong*, LI Chang-tian, YANG Yu, BAILin, WEI Lian, WANG Jing

    *Jilin Agricultural University，Changchun,Jilin 130000,China

    Abstract:   Objective  To establish a rapid, sensitive detection method for detecting RiftValley Fever virus (RVFV) by real-time fluorescence quantitativeRCR.    Methods   The primers and Taqmanprobe on the L segment conserved sequence of RVFV through alignmentwas designed. The real-time fluorescence quantitative PCRreactioning system was designed.   Results  Using a synthetic plasmid DNA as a positive control, thesensitivity with a specific real-time PCR method was 37 copies/μlfor RVFV, and without cross-reaction for the detection Japaneseencephalitis virus, Yellow fever virus, Dengue virus, Chikungunyavirus.   Conclusion   This assay shows to besensitive, specificity and real-time detectioning, which has goodprospects of application for disease prevention of infectiousdiseases at ports.

    Key words:   Real-timePCR；Detection；Rift Valley fever

    《中国国境卫生检疫杂志》2012年6月刊