狂犬病病毒荧光定量RT-PCR快速检测试剂盒的评价及应用

Evaluation and application of one step real-time TaqMan RT-PCR kit for detection of classic rabies virus

基于目前在我国人群和动物中流行的狂犬病病毒(RV)均属于基因Ⅰ型,在本室所建立的基因Ⅰ型RV荧光定量RT-PCR方法的基础上,组装了可以便捷使用的RV快速检测试剂盒。对4 577份临床样品的检测结果表明,该试剂盒的检测灵敏度达4.68个TCID50,与《狂犬病防治技术规范》规定的套式RT-PCR灵敏度相当,而且稳定性良好,可以冷冻保存8个月以上。所研制的荧光定量RT-PCR快速检测试剂盒具有操作简便、快速、特异性强、灵敏度高、重复性好、稳定性强等优点,既适合单个临床样品的确诊,又适合RV流行病学监测的大规模筛查。

Based on the phylogentic analysis of the current rabies virus（RV） strains in China,it showed that all RVs in our country belong to lyssavirus genotype Ⅰ.In our laboratory,the one-step real-time RT-PCR（qRT-PCR） has been established to detect the rabies virus,and then all components and reaction parameters were optimized to develop a real-time TaqMan RT-PCR kit.The developed real-time RT-PCR kit could detect 4.68 TCID50.It＇s sensitivity is equal with Nested RT-PCR,and it could be stored over eight months at-20℃.It was demonstrated through detection of 4 577 clinical samples that the developed kit was the simple,sensitive,specific,accurate method for large scale of epidemiological surveillance and the rapid detection of rabies virus.