Development of two TaqMan real-time reverse transcription-PCR assays for the detection of severe acute respiratory syndrome coronavirus-2  

作　　者：Yiwei Liu Yingying Wang Xinming Wang Yan Xiao Lan Chen Li Guo Jianguo Li Lili Ren Jianwei Wang 

机构地区：[1]National Health Commission Key Laboratory of Systems Biology of Pathogens and Christophe Mérieux Laboratory,Institute of Pathogen Biology,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100730,China [2]Institutes of Biomedical Sciences,Shanxi University,Taiyuan 030006,China [3]Key Laboratory of Respiratory Disease Pathogenomics,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100730,China

出　　处：《Biosafety and Health》2020年第4期232-237,共6页生物安全与健康（英文）

基　　金：funded in part by the Project from the Ministry of Science and Technology in China(2020YFC0841200);the National Major Science&Technology Project for Control and Prevention of Major Infectious Diseases in China(2017ZX10103004);the Chinese Academy of Medical Sciences(CAMS)Innovation Fund for Medical Sciences(2020HY320001);the non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences(2019PT310029);the Chinese Academy of Medical Sciences(CAMS)Innovation Fund for Medical Sciences(2016-I2M-1-014).

摘　　要：The outbreak of coronavirus disease 2019(COVID-19)in Wuhan,China,was caused by a novel coronavirus(CoV),named severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).The rapid detection of viral nucleic acids is critical for the early identification of infected cases.We have developed two TaqMan real-time reverse transcription-PCR assays to detect SARS-CoV-2.The designed primers target the nucleocapsid(N)and open reading frame(ORF)1b gene regions,where the probes discriminate SARS-CoV-2 from other human and animal CoVs.The sensitivities are one genomic copy per reaction for theN gene assay and ten copies for the ORF 1b gene assay.The overall linear detection ranges are 1–10^(6)and 10–10^(6)copies per reaction for the N gene assay and the ORF 1b gene assay,respectively.Surveillance of 23 suspected COVID-19 patients demonstrated that SARS-CoV-2 could be detected from 100%(23/23)and 62.5%(16/23)of clinical specimens by the N gene assay and the ORF 1b gene assay,respectively.All of the samples not detected by the ORF 1b gene assay were throat swabs,indicating a lower viral load in the upper respiratory tract and the relatively lower sensitivity of the ORF 1b gene assay.The assays developed in the present study offer alternative diagnostic tests for COVID-19.

关 键 词：SARS-CoV-2 Real-time RT-PCR NUCLEOCAPSID Open reading frame 1b 

分 类 号：R563.1[医药卫生—呼吸系统] R440[医药卫生—内科学]