Construction of a dual-model aptasensor based on G-quadruplexes generated via rolling circle amplification for visual/sensitive detection of kanamycin

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.