Selection and characterization of a novel DNA aptamer for label-free fluorescence biosensing of ochratoxin A.
Selection and characterization of a novel DNA aptamer for label-free fluorescence biosensing of ochratoxin A.
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DOI：
10.3390/toxins6082435
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发表时间：
2014-08-15
期刊：
影响因子：
4.2
通讯作者：
DeRosa MC
中科院分区：
文献类型：
--
作者：
McKeague M;Velu R;Hill K;Bardóczy V;Mészáros T;DeRosa MC
核酸适体正在成为食品安全监测的有用分子识别工具。然而，实践和技术挑战限制了可并入新型感测方案的可用适体探针的数量和多样性。这项工作描述了与重要食品污染物赭曲霉毒素A（OTA）结合的新型DNA适体的选择。在15轮体外选择后，分析序列的OTA结合。与类似的食品掺假物相比，两种分离的适体表现出对这种真菌毒素的高亲和力结合和选择性。将这些序列以及截短的适体（结合所需的最小序列）并入基于SYBR®绿色I荧光的OTA生物传感方案中。该无标记检测平台能够快速、选择性和灵敏地进行OTA定量，检测限为9 nM，线性定量高达100 nM。
Nucleic acid aptamers are emerging as useful molecular recognition tools for food safety monitoring. However, practical and technical challenges limit the number and diversity of available aptamer probes that can be incorporated into novel sensing schemes. This work describes the selection of novel DNA aptamers that bind to the important food contaminant ochratoxin A (OTA). Following 15 rounds of in vitro selection, sequences were analyzed for OTA binding. Two of the isolated aptamers demonstrated high affinity binding and selectivity to this mycotoxin compared to similar food adulterants. These sequences, as well as a truncated aptamer (minimal sequence required for binding), were incorporated into a SYBR® Green I fluorescence-based OTA biosensing scheme. This label-free detection platform is capable of rapid, selective, and sensitive OTA quantification with a limit of detection of 9 nM and linear quantification up to 100 nM.