Analytical Sciences

Fluorometric Aptasensor for Determination of Escherichia coli O157:H7 by FRET Effect between Aminated Carbon Quantum Dots and Graphene Oxide

Tao PAN, Xueling SHAN, Ding JIANG, Lu QI, Wenchang WANG, and Zhidong CHEN

Jiangsu Key Laboratory of Advanced Catalytic Materials and Technology, School of Petrochemical Engineering, Changzhou University, Changzhou 213164, China

A fluorometric aptasensor based on Escherichia coli O157:H7 (E. coli O157:H7) aptamer labeled aminated carbon quantum dots (NH₂-CQDs) and graphene oxide (GO) for the determination of E. coli O157:H7 was developed. In this research, carboxyl group (–COOH) terminated E. coli O157:H7 aptamer was steadily labeled to NH₂-CQDs by amidation reaction, and played the role of energy donor and was responsible for chemical recognition. Correspondingly, GO served as an energy acceptor. The introduction of NH₂-CQDs not only made the aptamer bond stably through covalent bond, but also significantly enhanced the fluorescence intensity compared with general CQDs. The NH₂-CQDs-aptamer is adsorbed on the surface of GO through π-π stacking and hydrophobic interaction. The fluorescence of NH₂-CQDs-aptamer was quenched via fluorescence resonance energy transfer (FRET) between NH₂-CQDs and GO. After adding E. coli O157:H7, the specific binding affinity between NH₂-CQDs-aptamer and E. coli O157:H7 lead to desorption of NH₂-CQDs-aptamer from GO, and recovery of the fluorescence intensity of NH₂-CQDs-aptamer. Under the optimal conditions, the increased fluorescence intensity showed a good linear relationship to concentrations of E. coli O157:H7 in the range 10² – 10⁷ cells/mL, with a detection limit of 89 cells/mL. Furthermore, the developed method was successfully applied to the determination of E. coli O157:H7 in commercial milk samples.

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