裂谷热病毒荧光定量PCR检测方法的建立  

Real-time PCR method for rapid detection of Rift Valley Fever virus

作　　者：杨志红[1,2] 李长田[1] 杨宇[2] 白琳[2] 魏莲[2] 王静[2] 

机构地区：[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

出　　处：《中国国境卫生检疫杂志》2012年第3期145-148,共4页Chinese Journal of Frontier Health and Quarantine

基　　金：国家自然科学基金(30900053);国家质检总局科研基金项目(2010IK211)

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。Objective To establish a rapid, sensitive detection method for detecting Rift Valley Fever virus （RVFV） by real-time fluorescence quantitative RCR. Methods The primers and Taqman probe on the L segment conserved sequence of RVFV through alignment was designed. The real-time fluorescence quantitative PCR reactioning system was designed. Results Using a synthetic plasmid DNA as a positive control, the sensitivity with a specific real-time PCR method was 37 copies/ul for RVFV, and without cross-reaction for the detection Japanese encephalitis virus, Yellow fever virus, Dengue virus, Chikungunya virus. Conclusion This assay shows to be sensitive, specificity and real-time detectioning, which has good prospects of application for disease prevention of infectious diseases at ports.

关 键 词：荧光定量PCR 检测 裂谷热 

分 类 号：R535[医药卫生—内科学]