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Simultaneous electrochemical determination of ochratoxin A and fumonisin B1 with an aptasensor based on the use of a Y-shaped DNA structure on gold nanorods

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Abstract

A complementary DNA (cDNA) was designed to simultaneously hybridize with the ochratoxin A (OTA) aptamer and the fumonisin B1 (FB1) aptamer to form a unique Y-shaped DNA structure and to achieve simultaneous detection. Gold nanorods (AuNRs) were used to immobilize thionine (Th), thiolated ferrocene (Fc), thiolated OTA aptamer (Apt1), and thiolated FB1 aptamer (Apt2), to form an amplified signal element and a recognition element. The Apt1-AuNRs-Th complex and the Apt2-AuNRs-Fc complex hybridize with cDNA to form a unique Y-DNA structure on a gold electrode. This produces two initial electrochemical signals [with 177 μΑ cm−2 near −0.2 V, and 3121 μΑ cm−2 near +0.46 V (vs. Ag/AgCl)] by differential pulse voltammetry. Upon addition of 0.1 ng mL−1 OTA and 0.1 ng mL−1 FB1, the aptamers bind the two toxins. This results in the release of Apt1-AuNRs-Th and Apt2-AuNRs-Fc, so the peak currents densities decrease to 115 μΑ cm−2 and 209 μΑ cm−2. The assay allows simultaneous determination of OTA and FB1 in the 1.0 pg·mL−1 to 100 ng·mL−1 concentration ranges, with LODs of 0.47 and 0.26 pg·mL−1. The assay is reproducible, stable and specific. It was applied to the determination of OTA and FB1 in spiked beer, with recoveries between 89.0% and 102.0%.

Schematic representation of OTA and FB1 detection based on Apt2-AuNRs-Fc/Apt1-AuNRs-Th/cDNA/AuE. (AuNRs: Gold nanorods; Th: thionine; Fc: ferrocene; SH: thiol; BSA: Bovine serum albumin; cDNA: Complementary DNA; Apt1: Aptamer1; Apt2: Aptamer2; OTA: Ochratoxin A; FB1: Fumonisin B1).

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Acknowledgments

This study was funded by the National Key Research and Development Program of China (2018YFC1602800),the Natural Science Foundation of Henan Province (182300410188), the Fundamental Research Funds for the Henan Provincial Colleges and Universities in Henan University of Technology (2016RCJH04), and Key Scientific and Technological Project of Henan Province (192102310255).

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Authors and Affiliations

  1. College of Food Science and Technology, Henan Key Laboratory of Cereal and Oil Food Safety Inspection and Control, Henan University of Technology, No.100 Lianhua Street, Zhengzhou, Henan Province, 450001, People’s Republic of China

    Min Wei, Lingkun Xin & Shuo Feng

  2. College of Chemistry and Chemical Engineering, Henan University, Jin Ming Avenue, Kaifeng, Henan Province, 475004, People’s Republic of China

    Yong Liu

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  1. Min Wei
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Wei, M., Xin, L., Feng, S. et al. Simultaneous electrochemical determination of ochratoxin A and fumonisin B1 with an aptasensor based on the use of a Y-shaped DNA structure on gold nanorods. Microchim Acta 187, 102 (2020). https://doi.org/10.1007/s00604-019-4089-y

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