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<p><i>Fusarium temperatum</i> (<i>F. temperatum</i>) is a fungus whose infection can cause various diseases in maize plants, leading to premature death, and <i>F. temperatum</i> mycotoxin poses a serious threat to human and animal health. Rapid and early on-site detection of <i>F. temperatum</i> infection facilitates the prevention of disease progression, which is an unmet need. In this study, a droplet digital CRISPR-Cas12a-based platform (DD-Cas), combined with a rapid extraction procedure, was developed for amplification-free on-site detection of <i>F. temperatum</i>. The DD-Cas assay can rapidly detect <i>F. temperatum</i> genomic DNA in infected maize samples with high sensitivity (10<sup>2</sup> CFU/mL) and specificity. Furthermore, we developed a smartphone-based fluorescence microscope integrating the heating module, that could accurately detect infected samples within 30&#xa0;min, enabling low-cost point-of-care testing (POCT). This platform can avoid cross-contamination and amplification bias, thus having great potential for on field detection of pathogenic bacteria in agriculture.</p> Graphical Abstract <p></p>

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Integrated on-site detection of Fusarium temperatum based on a droplet digital CRISPR-based platform

  • 期刊论文

  • 发表日期：

    2026年3月2日

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  • Yingchun Li,
  • Zequan Ye,
  • Chenfei Zhao,
  • Yunyun Tan,
  • Jianai Chen,
  • Zhina Wu,
  • Yaqin Zhang,
  • Hangyu Guo,
  • Yuan Cheng,
  • Rui Wang,
  • Jiasi Wang,
  • Di Wang

摘要

Fusarium temperatum (F. temperatum) is a fungus whose infection can cause various diseases in maize plants, leading to premature death, and F. temperatum mycotoxin poses a serious threat to human and animal health. Rapid and early on-site detection of F. temperatum infection facilitates the prevention of disease progression, which is an unmet need. In this study, a droplet digital CRISPR-Cas12a-based platform (DD-Cas), combined with a rapid extraction procedure, was developed for amplification-free on-site detection of F. temperatum. The DD-Cas assay can rapidly detect F. temperatum genomic DNA in infected maize samples with high sensitivity (102 CFU/mL) and specificity. Furthermore, we developed a smartphone-based fluorescence microscope integrating the heating module, that could accurately detect infected samples within 30 min, enabling low-cost point-of-care testing (POCT). This platform can avoid cross-contamination and amplification bias, thus having great potential for on field detection of pathogenic bacteria in agriculture.

Graphical Abstract