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Competitive fluorometric assay for the food toxin T-2 by using DNA-modified silver nanoclusters, aptamer-modified magnetic beads, and exponential isothermal amplification

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Abstract

The authors describe an aptamer based assay for the mycotoxin T-2. The method is making use of exponential isothermal amplification reaction (EXPAR) and fluorescent silver nanoclusters (AgNCs). Free T-2 and cDNA (which is a DNA that is partially complementary to the aptamer) compete for binding to aptamer-modified magnetic beads. The cDNA collected by magnetic separation can be used as a primer to trigger EXPAR to obtain ssDNA. The C-base-rich ssDNA binds and reduces Ag(I) ion to form fluorescent AgNCs. Fluorescence is measured at excitation/emission wavelengths of 480/525 nm. T-2 can be detected by fluorometry with a detection limit as low as 30 fg·mL−1. The method was applied to analyse spiked oat and corn, and the average recoveries ranged from 97.3 to 102.3% and from 95.9 to 107.5%, respectively. The results were in good agreement with data of the commercial ELISA kit. The assay is highly sensitive, has a wide analytical range, good specificity and reliable operation. It provides a promising alternative for the standard method for quantitative detection of T-2.

Schematic presentation of fluorometric assay for T-2 based on aptamer-functionalized magnetic beads exponential, isothermal amplification reaction (EXPAR) and fluorescent silver nanoclusters (AgNCs).

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Acknowledgements

This work was supported by the National Key Research and Development Program of China (No. 2017YFC1200903, 2017YFF0108403, 2017YFC1601205), the National Natural Science Foundation of China (No.81703229, 31701711).

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Author notes

  1. Man Zhang and Yu Wang contributed equally to this work.

Authors and Affiliations

  1. Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety, Tianjin Institute of Environment and Operational Medicine, Tianjin, 300050, People’s Republic of China

    Man Zhang, Yu Wang, Shuai Yuan, Xuan Sun, Bingyang Huo, Jialei Bai, Yuan Peng, Baoan Ning & Zhixian Gao

  2. School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai, 200093, People’s Republic of China

    Man Zhang & Baolin Liu

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  1. Man Zhang
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Correspondence to Baolin Liu or Zhixian Gao.

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Zhang, M., Wang, Y., Yuan, S. et al. Competitive fluorometric assay for the food toxin T-2 by using DNA-modified silver nanoclusters, aptamer-modified magnetic beads, and exponential isothermal amplification. Microchim Acta 186, 219 (2019). https://doi.org/10.1007/s00604-019-3322-z

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