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A novel aptasensor based on DNA hydrogel for sensitive visual detection of ochratoxin A

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Abstract

A novel visual detection mode is proposed to improve the detection sensitivity for the determination of ochratoxin A (OTA). The mode is based on aptamer recognition and the signal amplification of rolling circle amplification (RCA) products self-assembled DNA hydrogel. Moreover, gold nanoparticles (AuNPs) were directly assembled inside the DNA hydrogel by adjusting the padlock probe sequences to achieve a stronger binding force between the DNA hydrogel and AuNPs; this avoids the need for modification of AuNPs with DNA sequences. In the presence of OTA, DNA hydrogel is formed. With higher concentrations of OTA, a larger amount of DNA hydrogel is formed. When AuNPs are added to the DNA hydrogel, AuNPs can be enclosed inside the DNA hydrogel. With more DNA hydrogel, there is less AuNPs in the supernatant. Thus, the absorbance of the supernatant is anti-correlated with the concentration of OTA. After optimization of the experimental conditions, the change in the absorbance of the supernatant was linearly correlated with the concentration of OTA, in the range 0.05 to 10 ng/mL; the limit of detection was 0.005 ng/mL. The good specificity of the developed biosensor was confirmed in the presence of other mycotoxins that are coexistent with or analogues of OTA. By comparing the developed method with the ELISA method, the accuracy and stability of this new method were also verified, with good performance obtained in real samples.

Graphical abstract

Diagram of the principle of the colorimetric aptasensor for OTA detection based on rolling circle amplification product self-assembled DNA hydrogel.

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Funding

This work was partly supported by the Science and Technology Innovation Action Plan of Shanghai (20392001600), the Natural Science Foundation of Shanghai (18ZR1409500), and the Technology R&D Program of Suzhou (SNG2018044).

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Authors and Affiliations

  1. School of Medical Instrument and Food Engineering, Shanghai Engineering Research Center for Food Rapid Detection, University of Shanghai for Science and Technology, Shanghai, 200093, People’s Republic of China

    Liling Hao & Fei Xu

  2. Samueli School of Engineering, University of California Los Angeles, Los Angeles, 90026, USA

    XuTao Liu

  3. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, People’s Republic of China

    Sunjie Xu & Faliang An

  4. School of Chemistry and Life Sciences, Suzhou University of Science and Technology, Suzhou, 215009, People’s Republic of China

    Huajie Gu

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  1. Liling Hao
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  2. XuTao Liu
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  3. Sunjie Xu
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  4. Faliang An
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  5. Huajie Gu
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  6. Fei Xu
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Correspondence to Fei Xu.

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Hao, L., Liu, X., Xu, S. et al. A novel aptasensor based on DNA hydrogel for sensitive visual detection of ochratoxin A. Microchim Acta 188, 395 (2021). https://doi.org/10.1007/s00604-021-05000-y

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  • DOI: https://doi.org/10.1007/s00604-021-05000-y

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