?Aptamer-locker? DNA coupling with CRISPR/Cas12a-guided biosensing for high-efficiency melamine analysis

文献类型： 外文期刊

第一作者： Qiao, Bin

作者： Qiao, Bin;Xu, Jiakun;Yin, Wenhao;Xin, Wanmeng;Ma, Lixin;Qiao, Jie;Liu, Yi;Yin, Wenhao;Xin, Wanmeng;Ma, Lixin;Qiao, Jie;Liu, Yi

作者机构：

关键词： Aptamer; CRISPR; Cas12a; Melamine; Biosensor; On-site test

期刊名称：BIOSENSORS & BIOELECTRONICS （ 影响因子：10.257； 五年影响因子：8.669 ）

ISSN： 0956-5663

年卷期： 2021 年 183 卷

页码：

收录情况： SCI

摘要： Herein, we report a method that combined ?aptamer-locker? DNA with CRISPR/Cas12a-based biosensing for sensitive and rapid melamine analysis. Three strategies were harnessed for designing the DNA sensors that were well characterized by circular dichroism (CD) spectroscopy and isothermal titration calorimetry (ITC) in the absence and presence of melamine. The detection parameters were optimized to achieve good analytic performance. As a result, a limit of detection (LOD) as low as 38 nM was achieved, which is below the threshold (1.0 mg/kg) of allowable melamine in infant milk products. In addition, the sensors show high selectivity for melamine against other analogues such as cyanuric acid, ammeline and ammelide. Moreover, our method was effective for rapid melamine analysis in whole milk samples, with or without sample pretreatment, in less than 20 min. Adopting a commercially available portable fluorimeter, on-site analysis of melamine in milk was accomplished. The strategies demonstrated here can expand to detect other non-nucleic-acid targets by simply replacing the aptamers.

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