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Fluorescence method for kanamycin detection based on the conversion of G-triplex and G-quadruplex

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Abstract

A versatile fluorescence scaffold was constructed by connecting a G-triplex sequence (G31) with G-rich DNA (aptamer of kanamycin) and using thioflavin T (ThT) as the fluorescent molecule. With the assistance of an aptamer, the G-quadruplex DNA structure was fabricated using G31 as three strands and the aptamer as the fourth strand. Due to the parallel planar morphology of the final products, which was favorable for ThT binding and which restricted the rotation of the aromatic rings of ThT, the fluorescence signal intensity of ThT was significantly enhanced. Because of the specific interaction of aptamer and kanamycin, in addition to the greater ability for kanamycin to bind with G-triplex than ThT, the conformation of G-quadruplex DNA was changed; in addition, ThT was dissociated from the aptamer-G31, and therefore a ‘turn-on’ to ‘turn-off’ detection principle was applied for kanamycin detection, which yielded reasonable sensitivity and selectivity. The detection range was 50–2000 nM, with a limit of detection of 1.05 nM. Our proposed method was thus successfully applied for kanamycin determination in pork, chicken, and beef samples, and satisfactory results were obtained.

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Acknowledgements

This study was funded by the National Natural Science Foundation of China (21305032), China Postdoctoral Science Foundation (2014M551522) and Jiangsu Planned Projects for Postdoctoral Research Funds (1402073B).

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Authors and Affiliations

  1. College of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, P. R. China

    Chengke Wang & Jiangyu Li

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  1. Chengke Wang
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  2. Jiangyu Li
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C. Wang contributed to the design of the experiments, data analysis and manuscript preparation. J. Li performed the experiments.

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Correspondence to Chengke Wang.

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Wang, C., Li, J. Fluorescence method for kanamycin detection based on the conversion of G-triplex and G-quadruplex. Anal Bioanal Chem 413, 7073–7080 (2021). https://doi.org/10.1007/s00216-021-03676-y

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  • DOI: https://doi.org/10.1007/s00216-021-03676-y

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