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Simultaneous detection of AFB1 and aflD gene by “Y” shaped aptamer fluorescent biosensor based on double quantum dots

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Abstract

The developed method for simultaneous detection of aflatoxin B1 (AFB1) and aflD genes can effectively monitor from the source and reduce the safety problems and economic losses caused by the production of aflatoxin, which can be of great significance for food safety regulations. In this paper, we constructed a sensitive and convenient fluorescent biosensor to detect AFB1 and aflD genes simultaneously based on fluorescence resonance energy transfer (FRET) between quantum dots (QDs) and a black hole quenching agent. A stable “Y” shaped aptasensor was employed as the detection platform and a double quantum dot labeled DNA fragment was utilized to be the sensing element in this work. When the targets of AFB1 and aflD genes were presented in the solution, the aptamer in the “Y” shaped probe is specifically recognized by the target. At this time, both Si-carbon quantum dots (Si-CDs) and CdTe QDs are far away from the BHQ1 and BHQ3 to recover the fluorescence. The linear range of the prepared fluorescence simultaneous detection method was as wide as 0.5–500 ng·mL−1 with detection lines of 0.64 ng·mL−1 for AFB1 and 0.5–500 nM with detection lines of 0.75 nM for aflD genes (3σ/k). This fabricated fluorescent biosensor was further validated in real rice flour and corn flour samples, which also achieved good results. The recoveries were calculated by comparing the known and found amounts of AFB1 which ranged from 88.4 to approximately 115.32% in the rice flour samples and 90.7 ~ 102.58% in the corn flour samples. The recoveries of aflD genes ranged from 84.32 to approximately 109.3% in the rice flour samples and 89.48 ~ 100.99% in the corn flour samples. Therefore, the proposed biosensor can significantly improve food safety and quality control through a simple, fast, and sensitive agricultural product monitoring and detection system.

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The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Funding

This work was supported by the National Natural Science Foundation of China (No. 31901799, 32001804), the China Postdoctoral Science Foundation (No. 2021M692370), the JUST Emerging Science and Technology Innovation Team Grant (No. 1182921902), the Jiangsu Dual Innovation Talent Program (No. 1184902001), and the Jiangsu University of Science and Technology Research Start-up Fund (No. 1182932001, 1182922001).

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Authors and Affiliations

  1. School of Grain Science and Technology, Jiangsu University of Science and Technology, Zhenjiang, 212100, Jiangsu Province, People’s Republic of China

    Yaqi Li, Xin Chen, Shuangfeng Peng, Dezhao Kong, Chang Liu, Qi Zhang & Qiaoqiao Shi

  2. School of Environmental Science and Engineering, Huazhong University of Science and Technology, Wuhan, 430074, Hubei Province, People’s Republic of China

    Yaqi Li & Yong Chen

  3. Advanced Technology Institute of Suzhou, Suzhou, 215123, Jiangsu Province, People’s Republic of China

    Yaqi Li

  4. School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang, 212100, Jiangsu Province, People’s Republic of China

    Qingyue Sun

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  1. Yaqi Li
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  2. Qingyue Sun
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  3. Xin Chen
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  4. Shuangfeng Peng
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  7. Qi Zhang
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  9. Yong Chen
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Contributions

Yaqi Li: funding acquisition, conceptualization, writing, review and editing. Qingyue Sun: conceptualization, investigation, writing—original draft. Xin Chen: the main strengths in the stage of article revision including synthesis and preparation new quantum dot probes, data analysis, and literature searching and organization. Shuangfeng Peng: data curation, formal analysis. Dezhao Kong: supervision. Chang Liu: validation. Qi Zhang and Qiaoqiao Shi: methodology. Yong Chen: supervision.

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Li, Y., Sun, Q., Chen, X. et al. Simultaneous detection of AFB1 and aflD gene by “Y” shaped aptamer fluorescent biosensor based on double quantum dots. Anal Bioanal Chem 416, 883–893 (2024). https://doi.org/10.1007/s00216-023-05074-y

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