Analytical comparisons of SARS-COV-2 detection by qRT-PCR and ddPCR with multiple primer/probe sets
ABSTRACTDifferent primers/probes sets have been developed all over the world for the nucleic acid detection of SARS-CoV-2 by quantitative real time polymerase chain reaction (qRT-PCR) as a standard method. In our recent study, we explored the feasibility of droplet digital PCR (ddPCR) for clinical S...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2020-01-01
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| Series: | Emerging Microbes and Infections |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/22221751.2020.1772679 |
| _version_ | 1797265892486152192 |
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| author | Xinjin Liu Jiangpeng Feng Qiuhan Zhang Dong Guo Lu Zhang Tao Suo Wenjia Hu Ming Guo Xin Wang Zhixiang Huang Yong Xiong Guozhong Chen Yu Chen Ke Lan |
| author_facet | Xinjin Liu Jiangpeng Feng Qiuhan Zhang Dong Guo Lu Zhang Tao Suo Wenjia Hu Ming Guo Xin Wang Zhixiang Huang Yong Xiong Guozhong Chen Yu Chen Ke Lan |
| author_sort | Xinjin Liu |
| collection | DOAJ |
| description | ABSTRACTDifferent primers/probes sets have been developed all over the world for the nucleic acid detection of SARS-CoV-2 by quantitative real time polymerase chain reaction (qRT-PCR) as a standard method. In our recent study, we explored the feasibility of droplet digital PCR (ddPCR) for clinical SARS-CoV-2 nucleic acid detection compared with qRT-PCR using the same primer/probe sets issued by Chinese Center for Disease Control and Prevention (CDC) targeting viral ORF1ab or N gene, which showed that ddPCR could largely minimize the false negatives reports resulted by qRT-PCR [Suo T, Liu X, Feng J, et al. ddPCR: a more sensitive and accurate tool for SARS-CoV-2 detection in low viral load specimens. medRxiv [Internet]. 2020;2020.02.29.20029439. Available from: https://medrxiv.org/content/early/2020/03/06/2020.02.29.20029439.abstract]. Here, we further stringently compared the performance of qRT-PCR and ddPCR for 8 primer/probe sets with the same clinical samples and conditions. Results showed that none of 8 primer/probe sets used in qRT-PCR could significantly distinguish true negatives and positives with low viral load (10−4 dilution). Moreover, false positive reports of qRT-PCR with UCDC-N1, N2 and CCDC-N primers/probes sets were observed. In contrast, ddPCR showed significantly better performance in general for low viral load samples compared to qRT-PCR. Remarkably, the background readouts of ddPCR are relatively lower, which could efficiently reduce the production of false positive reports. |
| first_indexed | 2024-03-07T17:24:20Z |
| format | Article |
| id | doaj.art-b96cc79001f9401ab1b6ac2a49651ee2 |
| institution | Directory Open Access Journal |
| issn | 2222-1751 |
| language | English |
| last_indexed | 2024-04-25T00:52:01Z |
| publishDate | 2020-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Emerging Microbes and Infections |
| spelling | doaj.art-b96cc79001f9401ab1b6ac2a49651ee22024-03-11T16:04:24ZengTaylor & Francis GroupEmerging Microbes and Infections2222-17512020-01-01911175117910.1080/22221751.2020.1772679Analytical comparisons of SARS-COV-2 detection by qRT-PCR and ddPCR with multiple primer/probe setsXinjin Liu0Jiangpeng Feng1Qiuhan Zhang2Dong Guo3Lu Zhang4Tao Suo5Wenjia Hu6Ming Guo7Xin Wang8Zhixiang Huang9Yong Xiong10Guozhong Chen11Yu Chen12Ke Lan13State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People’s Republic of ChinaState Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People’s Republic of ChinaState Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People’s Republic of ChinaState Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People’s Republic of ChinaState Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People’s Republic of ChinaState Key Laboratory of Virology, Renmin Hospital, Wuhan University, Wuhan, People’s Republic of ChinaDepartment of Infectious Disease, Zhongnan Hospital, Wuhan University, Wuhan, People’s Republic of ChinaState Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People’s Republic of ChinaState Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People’s Republic of ChinaState Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People’s Republic of ChinaDepartment of Infectious Disease, Zhongnan Hospital, Wuhan University, Wuhan, People’s Republic of ChinaState Key Laboratory of Virology, Renmin Hospital, Wuhan University, Wuhan, People’s Republic of ChinaState Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People’s Republic of ChinaState Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People’s Republic of ChinaABSTRACTDifferent primers/probes sets have been developed all over the world for the nucleic acid detection of SARS-CoV-2 by quantitative real time polymerase chain reaction (qRT-PCR) as a standard method. In our recent study, we explored the feasibility of droplet digital PCR (ddPCR) for clinical SARS-CoV-2 nucleic acid detection compared with qRT-PCR using the same primer/probe sets issued by Chinese Center for Disease Control and Prevention (CDC) targeting viral ORF1ab or N gene, which showed that ddPCR could largely minimize the false negatives reports resulted by qRT-PCR [Suo T, Liu X, Feng J, et al. ddPCR: a more sensitive and accurate tool for SARS-CoV-2 detection in low viral load specimens. medRxiv [Internet]. 2020;2020.02.29.20029439. Available from: https://medrxiv.org/content/early/2020/03/06/2020.02.29.20029439.abstract]. Here, we further stringently compared the performance of qRT-PCR and ddPCR for 8 primer/probe sets with the same clinical samples and conditions. Results showed that none of 8 primer/probe sets used in qRT-PCR could significantly distinguish true negatives and positives with low viral load (10−4 dilution). Moreover, false positive reports of qRT-PCR with UCDC-N1, N2 and CCDC-N primers/probes sets were observed. In contrast, ddPCR showed significantly better performance in general for low viral load samples compared to qRT-PCR. Remarkably, the background readouts of ddPCR are relatively lower, which could efficiently reduce the production of false positive reports.https://www.tandfonline.com/doi/10.1080/22221751.2020.1772679SARS-CoV-2diagnosisdigital PCRreal time PCRfalse positivefalse negative |
| spellingShingle | Xinjin Liu Jiangpeng Feng Qiuhan Zhang Dong Guo Lu Zhang Tao Suo Wenjia Hu Ming Guo Xin Wang Zhixiang Huang Yong Xiong Guozhong Chen Yu Chen Ke Lan Analytical comparisons of SARS-COV-2 detection by qRT-PCR and ddPCR with multiple primer/probe sets Emerging Microbes and Infections SARS-CoV-2 diagnosis digital PCR real time PCR false positive false negative |
| title | Analytical comparisons of SARS-COV-2 detection by qRT-PCR and ddPCR with multiple primer/probe sets |
| title_full | Analytical comparisons of SARS-COV-2 detection by qRT-PCR and ddPCR with multiple primer/probe sets |
| title_fullStr | Analytical comparisons of SARS-COV-2 detection by qRT-PCR and ddPCR with multiple primer/probe sets |
| title_full_unstemmed | Analytical comparisons of SARS-COV-2 detection by qRT-PCR and ddPCR with multiple primer/probe sets |
| title_short | Analytical comparisons of SARS-COV-2 detection by qRT-PCR and ddPCR with multiple primer/probe sets |
| title_sort | analytical comparisons of sars cov 2 detection by qrt pcr and ddpcr with multiple primer probe sets |
| topic | SARS-CoV-2 diagnosis digital PCR real time PCR false positive false negative |
| url | https://www.tandfonline.com/doi/10.1080/22221751.2020.1772679 |
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