DNAzyme-driven Cascade DNA Walker Fluorescent Biosensor for Highly Sensitive Detection of Kanamycin

doi:10.1007/s40242-025-5062-4

Chemical Research in Chinese Universities ›› 2025, Vol. 41 ›› Issue (4): 975-982.doi: 10.1007/s40242-025-5062-4

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DNAzyme-driven Cascade DNA Walker Fluorescent Biosensor for Highly Sensitive Detection of Kanamycin

NIE Saiyu, ZHANG Peng, ZHANG Qian DING Caifeng

Laboratory of Optic-electric Sensing and Analytical Chemistry for Life Science of Ministry of Education, Shandong Key Laboratory of Biochemical Analysis, Key Laboratory of Analytical Chemistry for Life Science in Universities of Shandong, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, P. R. China

Received:2025-04-14 Accepted:2025-05-06 Online:2025-08-01 Published:2025-07-24
Supported by:
This work was supported by the National Natural Science Foundation of China (Nos. 22474068, 22004078 and 22074074), the Youth Innovation Team Development Program of Shandong Higher Education Institutions, China (No. 2022KJC004), the Natural Science Foundation of Shandong Province, China (No. ZR2024QB063), and the Natural Science Foundation of Qingdao, China (No. 23-2-1-175-zyyd-jch).

Abstract

Abstract: The contamination of marine environments with antibiotics, such as kanamycin, poses a significant ecological threat due to its potential harmful effects on marine organisms. Conventional methods for detecting kanamycin, including high-performance liquid chromatography (HPLC) and enzyme-linked immunosorbent assays (ELISA), face challenges in terms of cost, complexity, and long detection times. This research aims to address these limitations by developing a novel, highly sensitive biosensing platform for kanamycin detection based on a DNAzyme-driven cascade reaction coupled with nanoparticle technology. The method utilizes aptamers for specific kanamycin recognition, which activates DNAzyme-1, triggering a downstream DNA walker system. This cascade reaction amplifies fluorescence signals, significantly enhancing sensitivity. The optimized platform demonstrated a wide detection range (0.05-3 nmol/L) and a low detection limit of 0.0327 nmol/L. This system showed excellent specificity for kanamycin, even in the presence of other antibiotics, and was successfully applied to detecting kanamycin in seawater samples with high recovery rates (94%-102.9%). This DNAzyme-based biosensor offers a promising approach for rapid, cost-effective, and high-sensitivity monitoring of antibiotic contamination in aquatic environments, with potential applications in environmental monitoring and ecological protection.

Key words: Kanamycin, Fluorescent biosensor, DNAzyme, DNA walker, Cascade amplification

NIE Saiyu, ZHANG Peng, ZHANG Qian DING Caifeng. DNAzyme-driven Cascade DNA Walker Fluorescent Biosensor for Highly Sensitive Detection of Kanamycin[J]. Chemical Research in Chinese Universities, 2025, 41(4): 975-982.

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DNAzyme-driven Cascade DNA Walker Fluorescent Biosensor for Highly Sensitive Detection of Kanamycin

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