TY - GEN
T1 - Rapid Virus Detection Using Recombinase Polymerase Amplification Assisted by Computational Amplicon-Complex Spectrum
AU - Yang, F.
AU - Su, Y.
AU - Li, F. G.
AU - Zhou, T. Q.
AU - Wang, X. S.
AU - Li, H.
AU - Zhang, S. L.
AU - Fu, R. X.
N1 - Publisher Copyright:
© The Author(s), under exclusive license to Springer Nature Switzerland AG 2024.
PY - 2024
Y1 - 2024
N2 - 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.
AB - 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.
KW - Computational spectrum
KW - Nucleic acid detection
KW - Recombinase polymerase amplification
KW - Virus detection
UR - http://www.scopus.com/inward/record.url?scp=85187795812&partnerID=8YFLogxK
U2 - 10.1007/978-3-031-51485-2_36
DO - 10.1007/978-3-031-51485-2_36
M3 - Conference contribution
AN - SCOPUS:85187795812
SN - 9783031514845
T3 - IFMBE Proceedings
SP - 335
EP - 342
BT - 12th Asian-Pacific Conference on Medical and Biological Engineering - Proceedings of APCMBE 2023-Volume 2
A2 - Wang, Guangzhi
A2 - Yao, Dezhong
A2 - Gu, Zhongze
A2 - Peng, Yi
A2 - Tong, Shanbao
A2 - Liu, Chengyu
PB - Springer Science and Business Media Deutschland GmbH
T2 - 12th Asian-Pacific Conference on Medical and Biological Engineering, APCMBE 2023
Y2 - 18 May 2023 through 21 May 2023
ER -