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A Novel Fluorescent Aptasensor Based on Dual-labeled DNA Nanostructure for Simultaneous Detection of Ochratoxin A and Aflatoxin B1

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Abstract

Based on DNA strand replacement reaction and aptamer-specific recognition, a simple dual-labeled DNA nanostructure is designed for the simultaneous detection of Ochratoxin A (OTA) and aflatoxin B1 (AFB1). C1 is labeled with Cy3 and Cy5, while C2 and C3 are labeled with BHQ2. The fluorescence intensity of DNA nanostructure composed of C1, C2 and C3 is weak because of fluorescence resonance energy transfer. When OTA Aptamer (OTA-Apt) and AFB1 Aptamer (AFB1-Apt) are added to the homogeneous system at the same time, C1 can be replaced with the help of toehold strand displacement, resulting in fluorescence enhancement. In the presence of both OTA and AFB1, the toehold strand displacement reaction is inhibited due to preferential binding between the target and their corresponding aptamers. The limit of detection of OTA was 0.007 ng/mL and that of AFB1 was 0.03 ng/mL. The recoveries of OTA and AFB1 were 96%-101% and 97%-101% in the corn sample, and 99%-101% and 92%-106% in the wine sample. Compared with other sensors, the preparation of this aptasensor needs simpler experimental steps and a shorter total-preparing time, confirming the convenient, rapid, and time-saving operation process.

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Data Availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Funding

This study was funded by the Key Scientific and Technological Project of Henan Province (212102310001), the Special Project for Collaborative Innovation of Zhengzhou (21ZZXTCX15), and the Innovative Funds Plan of Henan University of Technology (2021ZKCJ13).

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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, Zhengzhou, 450001, China

    Luyang Zhao, Xiujun Liang, Min Wei & Huali Jin

  2. College of Chemistry and Chemical Engineering, Henan University, Kaifeng, 475004, China

    Yong Liu

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  1. Luyang Zhao
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  4. Min Wei
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Contributions

Luyang Zhao: Investigation, Methodology, Validation and Writing-Original Draft. Xiujun Liang: Validation and Writing-Original Draft. Yong Liu: Supervision, Resources. Min Wei: Conceptualization, Methodology, Resources, Supervision and Funding acquisition. Huali Jin: Methodology, Resources, Supervision.

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Correspondence to Min Wei.

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Zhao, L., Liang, X., Liu, Y. et al. A Novel Fluorescent Aptasensor Based on Dual-labeled DNA Nanostructure for Simultaneous Detection of Ochratoxin A and Aflatoxin B1. J Fluoresc 33, 1641–1650 (2023). https://doi.org/10.1007/s10895-022-03071-5

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