A New Auto-RPA-Fluorescence Detection Platform for SARS-CoV-2

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/mu L 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.

Automated summary

The researchers developed a new auto-recombinase polymerase amplification (RPA)-fluorescence platform for the diagnosis of SARS-CoV-2. By optimizing primers and probes, the platform could detect SARS-CoV-2 nucleotides within 15 minutes, with high specificity and accuracy.