An exonuclease I-assisted quencher-free 2-aminopurine aptasensor based on a multipath paper-based device for ultrasensitive detection of kanamycin

文献类型： 外文期刊

第一作者： He, Xuan

作者： He, Xuan;Fu, Xiuli;Qi, Ji;Song, Dean

作者机构：

关键词： Paper-based device; 2-Aminopurine; Exonuclease I; Kanamycin; Fluorescence

期刊名称：MICROCHEMICAL JOURNAL （ 影响因子：4.9； 五年影响因子：4.5 ）

ISSN： 0026-265X

年卷期： 2024 年 203 卷

页码：

收录情况： SCI

摘要： A quencher-free multipath microfluidic paper-based analytical device (mu PAD) was constructed for ultrasensitive detection of kanamycin based on exonuclease I (Exo I)-assisted signal enhancement of 2-aminopurine (2-AP). Here, Exo I, a single-stranded DNA-specific nuclease, was introduced to fully liberate 2-AP mononucleotides to greatly enhance biosensing sensitivity. 2-AP, a fluorescent adenine analogue embedded in single-stranded DNA (ssDNA), was employed as the detection signal source. The fluorescence of 2-AP is strong in the mononucleotide state, while it can have low fluorescence and even no fluorescence in ssDNA and dsDNA, respectively. The 2-AP fluorescence probe included 2-AP DNA and kanamycin aptamer. When kanamycin was present, binding occurred between kanamycin and the aptamer, leading to ssDNA, which was further digested by Exo I. In this case, free 2AP mononucleotides were liberated, indicating strong fluorescence. In addition, the captured kanamycin was released for binding with the new aptamer, which resulted in the formation of a binding-hydrolysis-release cycle with the aid of Exo I. Under optimal conditions, this mu PAD exhibited sensitive and multipath detection of kanamycin at concentrations as low as 1.26 x 10-14 M with a wide range of 10- 13-10- 7 M. Furthermore, satisfactory results were achieved for analysing spiked kanamycin in milk and honey samples. This strategy is a very promising tool for monitoring antibiotics and evaluating the safety of animal-derived foods.

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