A Rapid and High-Sensitive Real-Time Reverse Transcription-Polymerase Chain Reaction Assay Used for the Detection of Severe Acute Respiratory Syndrome Coronavirus 2

Xiaomin Chen, Yan Xu, Jing Tian, Xueling Li, Hui Liang, Ang Gao, Zexi Liu, Dicheng Yang, Qi Shen, Daxiang Cui

Abstract

The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a public health emergency of international concern. Real-time reverse transcription-polymerase chain reaction (RT-PCR) is widely used as the gold standard method for the diagnosis of SARS-CoV-2 infection. However, the reliability of current real-time RT-PCR assays is questioned due to some false-negative reports. In this study, we improved the real-time RT-PCR method based on three target regions (ORF1ab, E, and N) of SARS-CoV-2. Results showed that real-time RT-PCR assays herein could complete detection within one hour after viral RNA preparation and had high sensitivity down to 5 copies of viral RNA. In addition, six clinical specimens were detected to evaluate the availability of this method. Among them, four samples were 3-plex SARS-CoV-2 positive and two were negative by real-time RT-PCR. The sensitivity was 100% (4/4), and specificity was 100% (2/2). These results demonstrate that we develop a rapid and high-sensitive real-time RT-PCR method for SARS-CoV-2 detection, which will be a powerful tool for COVID-19 identification and for monitoring suspected patients.

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Nano Biomedicine and Engineering.

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