Comparison of three TaqMan Real-Time Reverse Transcription-PCR assays in detecting SARS-CoV-2
Comparison of three TaqMan Real-Time Reverse Transcription-PCR assays in detecting SARS-CoV-2
Abstract Quick and accurate detection of SARS-CoV-2 is critical for COVID-19 control. Dozens of real-time reverse transcription PCR (qRT-PCR) assays have been developed to meet the urgent need of COVID-19 control. However, methodological comparisons among the developed qRT-PCR assays are limited. In the present study, we evaluated the sensitivity, specificity, amplification efficiency, and linear detection ranges of three qRT-PCR assays, including the assays developed by our group (IPBCAMS), and the assays recommended by WHO and China CDC (CCDC). The three qRT-PCR assays exhibited similar sensitivities, with the limit of detection (LOD) at about 10 copies per reaction (except the ORF 1b gene assay in CCDC assays with a LOD at about 100 copies per reaction). No cross reaction with other respiratory viruses were observed in all of the three qRT-PCR assays. Wide linear detection ranges from 106 to 101 copies per reaction and acceptable reproducibility were obtained. By using 25 clinical specimens, the N gene assay of IPBCAMS assays and CCDC assays performed better (with detection rates of 92% and 100%, respectively) than that of the WHO assays (with a detection rate of 60%), and the ORF 1b gene assay in IPBCAMS assays performed better (with a detection rate of 64%) than those of the WHO assays and the CCDC assays (with detection rates of 48% and 20%, respectively). In conclusion, the N gene assays of CCDC assays and IPBCAMS assays and the ORF 1b gene assay of IPBCAMS assays were recommended for qRT-PCR screening of SARS-CoV-2.
Wang Yingying、Wang Ying、Wang Geng、Li Jianguo、Xiao Yan、Ren Lili、Li Zhen、Wang Xinming
National Health Commission Key Laboratory of Systems Biology of Pathogens and Christophe M¨|rieux Laboratory, Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical CollegeNational Health Commission Key Laboratory of Systems Biology of Pathogens and Christophe M¨|rieux Laboratory, Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical CollegeNational Health Commission Key Laboratory of Systems Biology of Pathogens and Christophe M¨|rieux Laboratory, Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical CollegeInstitutes of Biomedical Sciences, Shanxi UniversityNational Health Commission Key Laboratory of Systems Biology of Pathogens and Christophe M¨|rieux Laboratory, Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical CollegeNational Health Commission Key Laboratory of Systems Biology of Pathogens and Christophe M¨|rieux Laboratory, Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College||Key Laboratory of Respiratory Disease Pathogenomics, Chinese Academy of Medical Sciences and Peking Union Medical CollegeInstitutes of Biomedical Sciences, Shanxi UniversityNational Health Commission Key Laboratory of Systems Biology of Pathogens and Christophe M¨|rieux Laboratory, Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College
医学研究方法基础医学微生物学
SARS-CoV-2qRT-PCRmethodological evaluationLimit of Detectionreproductivity clinical performance
Wang Yingying,Wang Ying,Wang Geng,Li Jianguo,Xiao Yan,Ren Lili,Li Zhen,Wang Xinming.Comparison of three TaqMan Real-Time Reverse Transcription-PCR assays in detecting SARS-CoV-2[EB/OL].(2025-03-28)[2025-08-09].https://www.biorxiv.org/content/10.1101/2020.07.06.189860.点此复制
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