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<p><i>Vibrio anguillarum</i> is a virulent pathogen responsible for severe diseases in mariculture. Its accurate on-site detection remains a challenge due to limitations in established methods, such as insufficient specificity, prolonged assay, and the requirement of non-movable instruments. We developed a visual platform for the rapid, specific, and sensitive detection of<i> V</i>. <i>anguillarum</i>. It integrates loop-mediated isothermal amplification (LAMP) with DNA-functionalized gold nanoparticles (DNA‒AuNPs); we termed it “nanoprobe-enhanced LAMP.” These DNA‒AuNP probes enabled the colorimetric discrimination of <i>empA</i> expression levels within 20&#xa0;min, while reducing the limit of detection to 1&#xa0;fg/μL level (equivalent to 89&#xa0;CFU/mL). More importantly, this technique demonstrated exceptional specificity in distinguishing <i>V. anguillarum</i> from other <i>Vibrio</i> spp. and foodborne bacteria via selective DNA recognition, outperforming conventional qPCR and fluorescence-based LAMP assays. We also exhibited the applicability of this method by detecting <i>V. anguillarum</i> in various tissues of infected turbots (<i>Scophthalmus maximus</i>), including liver, spleen, kidney, and skin; these results were consistent with the pathological findings. Given its simplicity, portability, and robustness—requiring only a constant temperature source—this approach is ideally suited for field-deployable diagnostics in resource-limited aquaculture settings.</p>

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Nanoprobe-enhanced loop-mediated isothermal amplification detection of Vibrio anguillarum in turbots (Scophthalmus maximus): toward advanced fish disease management

  • 期刊论文

  • 发表日期：

    2026年3月19日

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  • Feng Liu,
  • Menghan Lu,
  • Guanhua Xuan,
  • Kexin Jiang,
  • Xiaokun Wang,
  • Yonghua Hu,
  • Weiwei Zhang,
  • Jiin Jung,
  • Soon-Mi Shim,
  • Guoqing Wang

摘要

Vibrio anguillarum is a virulent pathogen responsible for severe diseases in mariculture. Its accurate on-site detection remains a challenge due to limitations in established methods, such as insufficient specificity, prolonged assay, and the requirement of non-movable instruments. We developed a visual platform for the rapid, specific, and sensitive detection of V. anguillarum. It integrates loop-mediated isothermal amplification (LAMP) with DNA-functionalized gold nanoparticles (DNA‒AuNPs); we termed it “nanoprobe-enhanced LAMP.” These DNA‒AuNP probes enabled the colorimetric discrimination of empA expression levels within 20 min, while reducing the limit of detection to 1 fg/μL level (equivalent to 89 CFU/mL). More importantly, this technique demonstrated exceptional specificity in distinguishing V. anguillarum from other Vibrio spp. and foodborne bacteria via selective DNA recognition, outperforming conventional qPCR and fluorescence-based LAMP assays. We also exhibited the applicability of this method by detecting V. anguillarum in various tissues of infected turbots (Scophthalmus maximus), including liver, spleen, kidney, and skin; these results were consistent with the pathological findings. Given its simplicity, portability, and robustness—requiring only a constant temperature source—this approach is ideally suited for field-deployable diagnostics in resource-limited aquaculture settings.