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Article

A Novel Fluorescent Aptasensor Based on Real-Time Fluorescence and Strand Displacement Amplification for the Detection of Ochratoxin A

by 1,2, 3, 1, 1, 4, 1 and 1,5,6,*
1
College of Food Science and Technology, Hebei Agricultural University, Baoding 071000, China
2
Hebei Province Feed Microorganism Technology Innovation Center, Baoding 071000, China
3
Department of Physical Education, Hebei Agricultural University, Baoding 071000, China
4
College of Science and Technology, Hebei Agricultural University, Cangzhou 061100, China
5
College of Life Sciences, Hebei Agricultural University, Baoding 071000, China
6
Hebei Key Laboratory of Analysis and Control of Zoonotic Pathogenic Microorganism, Hebei Agricultural University, Baoding 071001, China
*
Author to whom correspondence should be addressed.
Academic Editors: Francesco Arioli and Federica Di Cesare
Foods 2022, 11(16), 2443; https://doi.org/10.3390/foods11162443
Received: 4 July 2022 / Revised: 4 August 2022 / Accepted: 10 August 2022 / Published: 13 August 2022
It is urgently necessary to develop convenient, reliable, ultrasensitive and specific methods of ochratoxin A determination in food safety owing to its high toxicity. In the present study, an ultrasensitive and labeled-free fluorescent aptamer sensor combining real-time fluorescence with strand displacement amplification (SDA) was fabricated for the determination of OTA. In the presence of OTA, the OTA–aptamer combines with OTA, thus opening hairpins. Then, SDA primers specifically bind to the hairpin stem, which is used for subsequent amplification as a template. SDA amplification is initiated under the action of Bst DNA polymerase and nicking endonuclease. The amplified products (ssDNA) are dyed with SYBR Green II and detected with real-time fluorescence. The method has good linearity in the range of 0.01–50 ng mL−1, with the lowest limit of detection of 0.01 ng mL−1. Additionally, the fluorescent aptamer sensor shows outstanding specificity and reproducibility. Furthermore, the sensor shows excellent analytical performance in the artificial labeled detection of wheat and oat samples, with a recovery rate of 96.1~100%. The results suggest that the developed sensor has a promising potential application for the ultrasensitive detection of contaminants in food. View Full-Text
Keywords: ochratoxin A; strand displacement amplification (SDA); real-time fluorescence; aptasensor ochratoxin A; strand displacement amplification (SDA); real-time fluorescence; aptasensor
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MDPI and ACS Style

Guo, W.; Yang, H.; Zhang, Y.; Wu, H.; Lu, X.; Tan, J.; Zhang, W. A Novel Fluorescent Aptasensor Based on Real-Time Fluorescence and Strand Displacement Amplification for the Detection of Ochratoxin A. Foods 2022, 11, 2443. https://doi.org/10.3390/foods11162443

AMA Style

Guo W, Yang H, Zhang Y, Wu H, Lu X, Tan J, Zhang W. A Novel Fluorescent Aptasensor Based on Real-Time Fluorescence and Strand Displacement Amplification for the Detection of Ochratoxin A. Foods. 2022; 11(16):2443. https://doi.org/10.3390/foods11162443

Chicago/Turabian Style

Guo, Wei, Haoyu Yang, Yunzhe Zhang, Hao Wu, Xin Lu, Jianxin Tan, and Wei Zhang. 2022. "A Novel Fluorescent Aptasensor Based on Real-Time Fluorescence and Strand Displacement Amplification for the Detection of Ochratoxin A" Foods 11, no. 16: 2443. https://doi.org/10.3390/foods11162443

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