Organophosphorus pesticides detection using broad-specific single-stranded DNA based fluorescence polarization aptamer assay
文献类型： 外文期刊
作者： Zhang, Cunzheng 1 ; Wang, Li 2 ; Tu, Zhui 1 ; Sun, Xing 2 ; He, Qinghua 1 ; Lei, Zhaojing 2 ; Xu, Chongxin 3 ; Liu, Yuan; 1 ;
作者机构： 1.Nanchang Univ, State Key Lab Food Sci & Technol, Nanchang 330047, Peoples R China
2.Jiangsu Acad Agr Sci, Inst Food Safety, Minist Agr, Key Lab Control Technol & Standard Agroprod Safet, Nanjing 210014, Jiangsu, Peoples R China
3.Jiangsu Acad Agr Sci, Inst Food Safety, Minist Agr, Key Lab Control Technol & Standard Agroprod Safet, Nanjing 2
关键词： Organophosphorus pesticides;DNA aptamer;Molecular beacon
期刊名称：BIOSENSORS & BIOELECTRONICS （ 影响因子：10.618； 五年影响因子：9.323 ）
ISSN：
年卷期：
页码：
收录情况： SCI
摘要： An approach is developed to detect the organophosphorus pesticides via competitive binding to a recombinant broad-specificity DNA aptamer with a molecular beacon (MB), the binding of the MB to the aptamer results in the activation of a fluorescent signal, which can be measured for pesticide quantification. Aptamers selected via the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) were structurally modified and truncated to narrow down the binding region of the target, which indicated that loops of the aptamer contributed different functions for different chemical recognition. Thereafter, a variant fused by two different minimum functional structures, was clarified with broad specificity and increased affinity. Further molecular docking and molecular dynamics simulations was conducted to understand the molecular interaction between DNA structure and chemicals. 3D modeling revealed a hot spot area formed by 3 binding sites, forces including hydrogen bonds and van der Waals interactions appear to play a significant role in enabling and stabilizing the binding of chemicals. Finally, an engineered aptamer based approach for the detection of organophosphorus pesticides was successfully applied in a test using a real sample, the limit of quantification (LOQ) for phorate, profenofos, isocarbophos, and omethoate reached 19.2,13.4,17.2, and 23.4 nM (0.005 mg L~(-1)), respectively.
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