?Aptamer-locker? DNA coupling with CRISPR/Cas12a-guided biosensing for high-efficiency melamine analysis
文献类型： 外文期刊
作者： Qiao, Bin 1 ; Xu, Jiakun 2 ; Yin, Wenhao 3 ; Xin, Wanmeng 3 ; Ma, Lixin 3 ; Qiao, Jie 3 ; Liu, Yi 3 ;
作者机构： 1.Zhengzhou Univ, Dept Oral & Maxillofacial Surg, Affiliated Hosp 1, Zhengzhou, Peoples R China
2.Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, Key Lab Sustainable Dev Polar Fisheries,Pilot Nat, Minist Agr & Rural Affairs,Lab Marine Drugs & Byp, Beijing, Peoples R China
3.Hubei Univ, State Key Lab Biocatalysts & Enzyme Engn, Wuhan, Peoples R China
4.Hubei Univ, Sch Life Sci, Hubei Key Lab Ind Biotechnol, Wuhan, Peoples R China
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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