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Simultaneous Detection of Ochratoxin A and Aflatoxin B1 Based on Stable Tuning Fork-shaped DNA Fluorescent Aptasensor

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Abstract

Tuning fork, consisting of two fork arms and a fork handle, has a stable and rigid structure. Inspired by this structure, a tuning fork-shaped DNA (TF-DNA) fluorescence aptasensor was constructed to detect ochratoxin A (OTA) and aflatoxin B1 (AFB1). A TF-DNA double-stranded structure capable of attaching both OTA aptamer labeled with the FAM fluorescent group (FAM-Apt) and AFB1 aptamer labeled with the ROX fluorescent group (ROX-Apt) was designed and linked to magnetic beads. This TF-DNA double-stranded structure can provide a stable platform for dual-target detection. In the presence of OTA and AFB1, FAM-Apt and ROX-Apt preferentially bound to them and detached from the TF-DNA double-stranded structure. Dual-signal fluorescent probes were collected from the supernatant by magnetic separation, and achieved fluorescence enhancement at 520 nm and 607 nm, respectively. The linear ranges are 0.05 ng/mL to 100 ng/mL for OTA and 0.1 ng/mL to 100 ng/mL for AFB1, and the detection limits are 0.015 ng/mL and 0.045 ng/mL, respectively. The developed sensor has the advantages of simple and fast preparation, good specificity and reproducibility, which is promising for the simultaneous determination of multiple hazardous substances in food.

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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 Special Project for Collaborative Innovation of Zhengzhou (21ZZXTCX15), the Innovative Funds Plan of Henan University of Technology (2021ZKCJ13), National Key Research and Development Program of China (2023YFF1104600), and Joint Research Fund for science and technology R&D Projects of Henan Province (225200810066).

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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, People’s Republic of China

    Mengyao Wang, Zeping Lv & Min Wei

  2. School of Energy Science and Technology, Henan University, Kaifeng, 475004, People’s Republic of China

    Yong Liu

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  1. Mengyao Wang
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  2. Zeping Lv
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Contributions

Mengyao Wang: Experimentation, Writing-original draft, Formal analysis, Investigation. Zeping Lv: Conceptualization, Investigation, Writing-review & editing. Yong Liu: Supervision, Data analysis. Min Wei: Conceptualization, Supervision, Funding acquisition, Resources, Writing-review & editing.

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Correspondence to Min Wei.

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Wang, M., Lv, Z., Liu, Y. et al. Simultaneous Detection of Ochratoxin A and Aflatoxin B1 Based on Stable Tuning Fork-shaped DNA Fluorescent Aptasensor. J Fluoresc (2024). https://doi.org/10.1007/s10895-024-03668-y

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