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Comparison of three TaqMan Real-Time Reverse Transcription-PCR assays in detecting SARS-CoV-2

Yan Xiao, Zhen Li, Xinming Wang, Yingying Wang, Ying Wang, Geng Wang, Lili Ren, Jianguo Li
doi: https://doi.org/10.1101/2020.07.06.189860
Yan Xiao
1National 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, P. R. China
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Zhen Li
2Institutes of Biomedical Sciences, Shanxi University, Taiyuan, 030006, P. R. China
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Xinming Wang
1National 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, P. R. China
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Yingying Wang
1National 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, P. R. China
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Ying Wang
1National 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, P. R. China
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Geng Wang
1National 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, P. R. China
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Lili Ren
1National 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, P. R. China
3Key Laboratory of Respiratory Disease Pathogenomics, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100730, P. R. China
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  • For correspondence: lijg@sxu.edu.cn renliliipb@163.com
Jianguo Li
2Institutes of Biomedical Sciences, Shanxi University, Taiyuan, 030006, P. R. China
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  • For correspondence: lijg@sxu.edu.cn renliliipb@163.com
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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.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted July 06, 2020.
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Comparison of three TaqMan Real-Time Reverse Transcription-PCR assays in detecting SARS-CoV-2
Yan Xiao, Zhen Li, Xinming Wang, Yingying Wang, Ying Wang, Geng Wang, Lili Ren, Jianguo Li
bioRxiv 2020.07.06.189860; doi: https://doi.org/10.1101/2020.07.06.189860
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Comparison of three TaqMan Real-Time Reverse Transcription-PCR assays in detecting SARS-CoV-2
Yan Xiao, Zhen Li, Xinming Wang, Yingying Wang, Ying Wang, Geng Wang, Lili Ren, Jianguo Li
bioRxiv 2020.07.06.189860; doi: https://doi.org/10.1101/2020.07.06.189860

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