Rapid Detection of SARS-CoV-2 Using Reverse transcription RT-LAMP method

Weihua Yang, Xiaofei Dang, Qingxi Wang, Mingjie Xu, Qianqian Zhao, Yunying Zhou, Huailong Zhao, Li Wang, Yihui Xu, Jun Wang, Shuyi Han, Min Wang, Fenyan Pei, Yunshan Wan
2020 · doi:10.1101/2020.03.02.20030130
preprint OA: closed
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A rapid, one-step RT-LAMP assay detected SARS-CoV-2 RNA within 30 minutes with sensitivity similar to RT-PCR and 99% specificity in clinical specimens.

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Abstract

Corona Virus Disease 2019 (COVID-19) is a recently emerged life-threatening disease caused by SARS-CoV-2. Real-time fluorescent PCR (RT-PCR) is the clinical standard for SARS-CoV-2 nucleic acid detection. To detect SARS-CoV-2 early and control the disease spreading on time, a faster and more convenient method for SARS-CoV-2 nucleic acid detecting, RT-LAMP method (reverse transcription loop-mediated isothermal amplification) was developed. RNA reverse transcription and nucleic acid amplification were performed in one step at 63 °C isothermal conditions, and the results can be obtained within 30 minutes. ORF1ab gene, E gene and N gene were detected at the same time. ORF1ab gene was very specific and N gene was very sensitivity, so they can guarantee both sensitivity and specificity for SARS-CoV-2. The sensitivity of RT-LAMP assay is similar to RT-PCR, and specificity was 99% as detecting 208 clinical specimens. The RT-LAMP assay reported here has the advantages of rapid amplification, simple operation, and easy detection, which is useful for the rapid and reliable clinical diagnosis of SARS-CoV-2.

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