期刊
TALANTA
卷 116, 期 -, 页码 520-526出版社
ELSEVIER
DOI: 10.1016/j.talanta.2013.07.011
关键词
Ochratoxin A; Aptamer; G-quadruplex; DNAzyme; Calorimetric biosensor
资金
- National Natural Science Foundation of China [21175124]
- National Key Basic Research Development Project of China [2010CB933602]
- Mobility Grant from the Scientific Office of the French Embassy in China
We report a new label-free calorimetric aptasensor based on DNAzyme-aptamer conjugate for rapid and high-throughput detection of Ochratoxin A (OTA, a possible human carcinogen, group 2B) in wine. Two oligonucleotides were designed for this detection. One is N1 for biorecognition, which includes two adjacent sequences: the OTA-specific aptamer sequence and the horseradish peroxidase (HRP)-mimicking DNAzyme sequence. The other is a blocking DNA (B2), which is partially complementary to a part of the OTA aptamer and partially complementary to a part of the DNAzyme. The existence of OTA reduces the hybridization between N1 and B2. Thus, the activity of the non-hybridized DNAzyme is linearly correlated with the concentration of OTA up to 30 nM with a limit of detection of 4 nM (3 sigma). Meanwhile, a double liquid-liquid extraction (LLE) method is accordingly developed to purify OTA from wine. Compared with the existing HPLC-FD or immunoassay methods, the proposed strategy presents the most appropriate balance between accuracy and facility, resulting in a considerable improvement of real-time quality control, and thereby, preventing chronic poisoning caused by OTA contained red wine. (C) 2013 Elsevier B.V. All rights reserved.
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