Rapid Virus Detection Using Recombinase Polymerase Amplification Assisted by Computational Amplicon-Complex Spectrum

The detection of respiratory viruses is crucial in the setting of the SARS-CoV-2 outbreak. Recombinase polymerase amplification technology speeds up nucleic acid detection compared to the conventional polymerase chain reaction, and requires no professional operations. The integration, cost, and convenience issues with recombinase polymerase amplification detection limits its wide application. In this study, we used the polar GelRed dye's computational absorption spectrum to probe the amplicon. After bonding with DNA, GelRed molecules can transform into polar electric dipoles attributing to the asymmetry structure. Following centrifugal vibration, electrostatic contact resulted in the precipitation of dipoles. The supernatant's absorbance spectra changed once the precipitation was removed. The remaining GelRed molecule in the amplified product can be assessed to determine its composition. Based on this principle, we confirmed the viability of the suggested method, and also concentrated the GelRed dye. Finally, we tested the effectiveness of this technique using the synthetic Influenza A template and primer. 10° copies/μL of the template was the lowest concentration that can be detected. It was linearly correlated with the template concentration logarithm. This technique has offered a workable, practical, and affordable detection strategy for the quick pathogen identification.