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Analytical comparisons of SARS-COV-2 detection by qRT-PCR and ddPCR with multiple primer/probe sets.
Liu, Xinjin; Feng, Jiangpeng; Zhang, Qiuhan; Guo, Dong; Zhang, Lu; Suo, Tao; Hu, Wenjia; Guo, Ming; Wang, Xin; Huang, Zhixiang; Xiong, Yong; Chen, Guozhong; Chen, Yu; Lan, Ke.
  • Liu X; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Feng J; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Zhang Q; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Guo D; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Zhang L; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Suo T; State Key Laboratory of Virology, Renmin Hospital, Wuhan University, Wuhan, People's Republic of China.
  • Hu W; Department of Infectious Disease, Zhongnan Hospital, Wuhan University, Wuhan, People's Republic of China.
  • Guo M; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Wang X; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Huang Z; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Xiong Y; Department of Infectious Disease, Zhongnan Hospital, Wuhan University, Wuhan, People's Republic of China.
  • Chen G; State Key Laboratory of Virology, Renmin Hospital, Wuhan University, Wuhan, People's Republic of China.
  • Chen Y; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
  • Lan K; State Key Laboratory of Virology, Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, People's Republic of China.
Emerg Microbes Infect ; 9(1): 1175-1179, 2020 Dec.
Article in English | MEDLINE | ID: covidwho-361278
ABSTRACT
Different 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.
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Full text: Available Collection: International databases Database: MEDLINE Type: Article Main subject: Pneumonia, Viral / Coronavirus Infections / Multiplex Polymerase Chain Reaction / Real-Time Polymerase Chain Reaction / Betacoronavirus Subject: Pneumonia, Viral / Coronavirus Infections / Multiplex Polymerase Chain Reaction / Real-Time Polymerase Chain Reaction / Betacoronavirus Type of study: Diagnostic study Language: English Journal: Emerg Microbes Infect Clinical aspect: Diagnosis Year: 2020

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Full text: Available Collection: International databases Database: MEDLINE Type: Article Main subject: Pneumonia, Viral / Coronavirus Infections / Multiplex Polymerase Chain Reaction / Real-Time Polymerase Chain Reaction / Betacoronavirus Subject: Pneumonia, Viral / Coronavirus Infections / Multiplex Polymerase Chain Reaction / Real-Time Polymerase Chain Reaction / Betacoronavirus Type of study: Diagnostic study Language: English Journal: Emerg Microbes Infect Clinical aspect: Diagnosis Year: 2020