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Updated: Sep 21, 2025

Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction
Published on: October 6, 2022
Construction of a dual-model aptasensor based on G-quadruplexes generated via rolling circle amplification for
Xiaolin Gao1, Zhicong Sun1, Xiaoyang Wang1
1School of Agricultural Engineering and Food Science, Shandong University of Technology, No. 12 Zhangzhou Road, Zibo 255049, Shandong Province, China.
Abstract:
A dual-model colorimetric and electrochemical aptasensor was designed using a large number of G-quadruplexes generated by rolling circle amplification (RCA). Specific binding between target and aptamer during RCA yielded large numbers of G-quadruplexes. A colorimetric sensor was fabricated based on the interaction between the G-quadruplex and hemin, which altered the 3,3',5,5'-Tetramethylbenzidine (TMB)-catalyzed color reaction and facilitated the visual and semi-quantitative detection of kanamycin. An electrochemical sensor was constructed based on the strong interaction between the G-quadruplex and the methylene blue electrical signal molecule. Combining nanocomposites multi-walled carbon nanotubes-chitosan/gold nanoparticles (MWCNTs-CS/AuNPs) and RCA realized double-amplified electrochemical signals. Under optimized conditions, a linear relationship was obtained as the logarithm of different concentrations of kanamycin (KAN). The colorimetric aptasensor had a linear range of 1 × 102 nM to 1 × 103 nM with a detection limit of 1.949 nM. The electrochemical aptasensor had wider a linear range from 1 × 10-3 nM to 2.5 × 103 nM and a lower detection limit of 0.333 pM. The sensor combined the advantages of simple colorimetric visualization with the ultra-precision of electrochemical methods. Aptasensor showed good specificity and prevented interference. Furthermore, the prepared dual-model aptasensor facilitated the practical monitoring of KAN in milk.

