新型冠状病毒双重荧光RT-PCR检测方法  

Multiplex Real-time PCR method for rapid detection of Novel Coronavirus

作　　者：杨宇[1] 刘丽娟[1] 赵婷婷[1] 李辉 王莎莎[1] 徐焕州[1] 杨鹏飞[1] 王旺[1] 孙肖红[1] 王静[1] 

机构地区：[1]中国检验检疫科学研究院卫生检疫研究所,北京100123 [2]上海辉睿生物科技有限公司

出　　处：《中国国境卫生检疫杂志》2012年第5期331-336,共6页Chinese Journal of Frontier Health and Quarantine

摘　　要：目的为快速检测新型冠状病毒,防控疫情的输入,建立一种快速的双重实时荧光RT-PCR检测方法。方法对WHO公布的单重实时荧光RT-PCR检测引物和探针进行重新设计和优化,建立双重荧光RT-PCR反应体系,分别采用羧基荧光素(FAM)和绿色荧光蛋白(VIC)荧光基团标记探针,实现双基因的同时检测。结果经优化的双重实时荧光RT-PCR方法有较好的灵敏度和特异性,对阳性对照质粒检测灵敏度为31拷贝/μl,检测健康和普通发热人员咽拭子以及流感病毒无交叉反应。结论建立了双重实时荧光RT-PCR快速检测方法,提高检测效率和准确度的同时降低了成本,可用于新型冠状病毒的快速应急检测。Objective To establish a rapid,sensitive method to detect Novel Coronavirus which are acute infections with high case fatality rates by multiplex real-time fluorescence quantitative RT- PCR.Methods Taqman probes labeled by FAM and VIC.The multiplex real-time quantitative RT-PCR assay was optimized based on the reported mono assay. The sensitivity was evaluated and influenza virus and human swab were using to examine the specificity.Results A specie real-time RT-PCR method was developed with the sensitivity of 31copies/｜xl for Novel Coronavirus,a synthetic plasmid DNA as a positive control,and no cross reaction was found with Influenza virus and human swab samples. Conclusion This multiple assay has good prospects of application for rapid test.

关 键 词：实时荧光PCR 新型冠状病毒 检测 交叉反应 

分 类 号：R183.4[医药卫生—流行病学]