期刊
BIOSENSORS & BIOELECTRONICS
卷 183, 期 -, 页码 -出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2021.113233
关键词
Aptamer; CRISPR; Cas12a; Melamine; Biosensor; On-site test
类别
资金
- National Natural Science Foundation of China [21907027, 22007030]
- China Postdoctoral Science Foundation [2019M662573]
- Natural Science Foundation of Hubei Province [2020CFB253]
A method combining "aptamer-locker" DNA with CRISPR/Cas12a-based biosensing was developed for sensitive and rapid melamine analysis, achieving good detection performance with a low limit of detection. The approach demonstrated high sensitivity, selectivity, and real-time analysis capabilities for detecting melamine in dairy products.
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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