Objective: The outbreak of COVID-19 caused by SARS-CoV-2 has led to a serious worldwide pandemic. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR)-based methods were recommended for routine detection of SARS-CoV-2 RNA. Because the reaction time and analytical sensitivity of qRT-PCR limits the diagnosis of SARS-CoV-2, development of a quick process of SARS-CoV-2 detection technology with high analytical sensitivity remains urgent. Methods: We combined isothermal amplification and fluorescence detection technology to develop a new auto-recombinase polymerase amplification (RPA)-fluorescence platform that could be used in the diagnosis of SARS-CoV-2. Results: By optimization of primers and probes, the RPA platform could detect SARS-CoV-2 nucleotides within 15 min. The limits of detection and specificity of the auto-RPA-fluorescence platform were 5 copies/µL and 100%, respectively. The accuracy of detection of the auto-RPA-fluorescence platform in the 16 positive samples was 100%. Conclusion: The RPA platform is a potential technology for the diagnosis of SARS-CoV-2 infection.
Bibliographical noteKAUST Repository Item: Exported on 2022-10-10
Acknowledgements: This work was sponsored by the Doctoral Fund of Jining No.1 People’s Hospital (2019004, 2020005), Technology Development Program of Shandong Province (2018WS469), China Postdoctoral Science Foundation (2020T130073ZX), and Natural Science Foundation of Shandong Province (NM ZR2020QH272), China.
We gratefully acknowledge the assistance of Professor Huabao Xiong and Professor Dongmei Shi with the writing of the manuscript.
ASJC Scopus subject areas
- Clinical Biochemistry
- Biochemistry, medical