Journal
FOOD ANALYTICAL METHODS
Volume 10, Issue 6, Pages 1982-1990Publisher
SPRINGER
DOI: 10.1007/s12161-016-0771-4
Keywords
Homogeneous electrochemical detection; Ochratoxin A; Recycling amplification
Categories
Funding
- Scientific Research Foundation for Returned Scholars, Ministry of Education of China [LXKQ201301]
- Fujian Spark Program [2016S0042]
- Science and Technology Program of Fujian, China [2016 N0017]
- Natural Science Foundation of China [31571779]
- High and New project of Fujian Marine Fisheries Department [[2015]20]
- Fujian Production and Study project of Provincial Science and Technology Hall, China [2016 N5006]
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A simple, fast, sensitive, and homogeneous electrochemical method has been developed for ochratoxin A (OTA) determination, which combines the advantage of the high selectivity of aptamer to OTA and high efficiency of exonuclease (Exo) III-assisted recycling amplification mechanism. The metastable hairpin probe (HP) was designed into three parts: regions I is the aptamer of OTA as the target recognition; regions II is poly[dA] sequence as extended DNA (E-DNA); regions III as the target DNA (T-DNA) is complementary to a part of the aptamer to stabilize the hairpin-sharp form of the HP in the absence of OTA. The DNA probe with a methylene blue tag at its 3' terminus (MB-DNA) was designed to be complementary with T-DNA. Introduction of OTA into the assay leads to the formation change of HP from hairpin shape to open form, thus facilitating the hybridization between T-DNA and MB-DNA. The electrochemical signal is amplified through continuous Exo III cleavage. Under the optimal conditions, the differential pulse voltammetric (DPV) response had a linear relationship with the logarithm of OTA concentration in the range of 0.001 similar to 0.5 ng mL(-1). In addition, the developed method has been successfully applied to detect OTA in wheat standard quality control sample. This homogeneous electrochemical sensor may have a potential prospect in detecting other molecules or proteins, which possesses the corresponding aptamer, through easily designing the hairpin probe.
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