Study on Bioluminescent Detection of the SARS-CoV-2 ORF1ab Gene by Coupling Isothermal RNA Reverse Transcription Amplification with a Digital PCR: A Novel Approach

The COVID-19 pandemic, which began in December 2019 and is caused by the SARS-CoV-2 virus, is a global public health emergency. We developed a more sensitive and improved method for detecting SARS-CoV-2 by combining bioluminescence in real-time (BART) and reverse-transcriptase loop-mediated amplification (RT-LAMP) protocols (RT-LAMP- BART), which was also compatible with a digital LAMP system (Rainsuit), which did not allow for real-time quantification but did allow absolute quantification with a spectrophotometer. We also developed a simulated digital RT-LAMP approach using this same principle to enlarge the overall reaction volume and achieve real-time detection with a limit of detection of 10 copies/mL, with further improvements in the overall dynamic range of this assay system being achieved through improving RNA availability in samples to ensure the target RNA present in reaction. This study’s innovative approach is critical for reliable detection and sequencing of numerous viruses.

Author (S) Details

Zhongjie Fei
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China.

Rongbin Wei
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China.

Chu Cheng
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China.

Pengfeng Xiao
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China.

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