期刊
ANALYTICAL BIOCHEMISTRY
卷 508, 期 -, 页码 58-64出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2016.05.024
关键词
Patulin; Aptamer; SELEX; Graphene oxide; Enzyme-chromogenic
资金
- National Science and Technology Support Program of China [2015BAD17B02-5]
- NSFC [31401576, 31401575]
- China Postdoctoral Science Foundation [2015M580402]
- Synergetic Innovation Center of Food Safety and quality control of Jiangsu Province
- S&T Support Program of Jiangsu Province [BE2013732]
- [JUSRP11547]
Patulin (PAT) is a kind of mycotoxin that has serious harmful impacts on both food quality and human health. A high-affinity ssDNA aptamer that specifically binds to patulin was generated using systemic evolution of ligands by exponential enrichment (SELEX) assisted by graphene oxide (GO). After 15 rounds of positive and negative selection, a highly enriched ssDNA pool was sequenced and the representative sequences were subjected to binding assays to evaluate their affinity and specificity. Of the eight aptamer candidates tested, the sequence PAT-11 bound to patulin with high affinity and excellent selectivity with a dissociation constant (K-d) of 21.83 +/- 5.022 nM. The selected aptamer, PAT-11, was subsequently used as a recognition element to develop a detection method for patulin based on an enzyme-chromogenic substrate system. The colorimetric aptasensor exhibited a linear range from 50 to 2500 pg mL(-1), and the limit of detection was found to be 48 pg mL-1. The results indicated that GO-SELEX technology was appropriate for the screening of aptamers against small-molecule toxins, offering a promising application for aptamer-based biosensors. (C) 2016 Elsevier Inc. All rights reserved.
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