目的建立一种快速、敏感检测克里米亚-刚果出血热病毒的实时荧光RT-PCR检测方法。方法分析克里米亚-刚果出血热病毒S基因片段核酸序列的保守性,设计实时荧光RT-PCR引物和探针,优化引物和探针浓度,确定最佳反应体系,并进行灵敏性、重复性、特异性检测。结果建立的克里米亚-刚果出血热病毒实时荧光RT-PCR方法的检测灵敏性为7.26×10~2 copies/μl,重复性好,与汉坦病毒等无交叉反应。结论建立的克里米亚-刚果出血热病毒实时荧光RT-PCR法能准确、快速地检测克里米亚-刚果出血热病毒。 Objective To establish a rapid and sensitive real-time fluorescence RT-PCR method for the detection of Crimean-Congo hemorrhagic fever virus. Methods The conservativeness of the nucleic acid sequence of S gene of Crimea-Congo hemorrhagic fever virus was analyzed. Real-time fluorescent RT-PCR primers and probes were designed, and the primers and probe concentrations were optimized to determine the optimal reaction system. Sensitivity and repeatability Sexual, specific testing. Results The detection sensitivity of the real-time RT-PCR method for Crimean-Congo haemorrhagic fever virus was 7.26 × 10 ~ 2 copies / μl with good repeatability and no cross-reaction with Hantavirus. Conclusions The Crimean-Congo hemorrhagic fever virus real-time fluorescence RT-PCR method can accurately and rapidly detect the Crimean-Congo hemorrhagic fever virus.