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Smartphone-based paper microfluidic competitive immunoassay for the detection of α-amanitin from mushrooms

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Abstract

α-Amanitin is often considered the most poisonous mushroom toxin produced by various mushroom species, which are hard to identify from edible, non-toxic mushrooms. Conventional detection methods require expensive and bulky equipment or fail to meet high analytical sensitivity. We developed a smartphone-based fluorescence microscope platform to detect α-amanitin from dry mushroom tissues. Antibody-nanoparticle conjugates were captured by immobilized antigen-hapten conjugates while competing with the free analytes in the sample. Captured fluorescent nanoparticles were excited at 460 nm and imaged at 500 nm. The pixel numbers of such nanoparticles in the test zone were counted, showing a decreasing trend with increasing analyte concentration. The detection method exhibited a low detection limit (1 pg/mL), high specificity, and selectivity, allowing us to utilize a simple rinsing for toxin extraction and avoiding the need for high-speed centrifugation. In addition, this assay’s short response time and portable features enable field detection of α-amanitin from amanitin-producing mushrooms.

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Acknowledgements

The authors are grateful for co-author Luisa W. Cheng’s contributions; she sadly passed away during the preparation of this manuscript. The authors acknowledge Kenneth E. Schackart III at the University of Arizona for his help on the initial discussion and initial assay optimization. The authors also acknowledge Lane E. Breshears, Brandon T. Nguyen, Patarajarin Akarapipad, and Katelyn Sosnowski, all at the University of Arizona, for their help in the design and fabrication of the smartphone-based fluorescence microscope.

Funding

The authors acknowledge the financial support from the National Institute of Food and Agriculture (NIFA), United States Department of Agriculture (USDA), provided through the Agriculture, Life and Veterinary Sciences, and Cooperative Extension at the University of Arizona. The authors also acknowledge the financial support from the Western Regional Research Center at the United States Department of Agriculture, Agricultural Research Service (USDA/ARS) at Albany, CA, USA.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, The University of Arizona, Tucson, AZ, USA

    Yan Liang & Jeong-Yeol Yoon

  2. Department of Biomedical Engineering, The University of Arizona, Tucson, AZ, USA

    Avory Zhou & Jeong-Yeol Yoon

  3. Foodborne Toxin Detection and Prevention Research Unit, Department of Agriculture, Agricultural Research Service, Western Regional Research Center, Albany, CA, USA

    Candace S. Bever & Luisa W. Cheng

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  1. Yan Liang
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  2. Avory Zhou
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  3. Candace S. Bever
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  4. Luisa W. Cheng
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  5. Jeong-Yeol Yoon
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Correspondence to Jeong-Yeol Yoon.

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Liang, Y., Zhou, A., Bever, C.S. et al. Smartphone-based paper microfluidic competitive immunoassay for the detection of α-amanitin from mushrooms. Microchim Acta 189, 322 (2022). https://doi.org/10.1007/s00604-022-05407-1

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