<p> The&#xa0;use of ultrasmall high-entropy alloys (HEAs, ≈5&#xa0;nm) with unique catalytic properties, integrating them into lateral flow immunoassay&#xa0;(LFIA) for the detection of <i>Staphylococcus aureus</i>&#xa0;is proposed.&#xa0;Under optimal conditions, the detection limits of the HEAs-based LFIA were 1.5 × 10<sup>3</sup>&#xa0;CFU/mL via direct coloration and 15&#xa0;CFU/mL using the DAB-enhanced catalytic signal, representing improvements of 10<sup>3</sup>-fold and 10<sup>5</sup>-fold, respectively. The HEAs demonstrated superior performance compared with traditional AuNPs, including enhanced sensitivity and high peroxidase activity (Km = 0.066&#xa0;M, Vmax = 1.132 × 10<sup>–6</sup>&#xa0;M&#xa0;min<sup>−1</sup>), as well as excellent specificity and repeatability, with relative standard deviation (RSD) of less than 10%. Moreover, the incubation time for HEAs functionalized with antibodies to bind <i>Staphylococcus aureus</i> was only 5&#xa0;min, compared with&#xa0;20&#xa0;min for functionalized AuNPs. The method also showed high specificity, effectively discriminating <i>Staphylococcus aureus</i> from other foodborne pathogens. The HEAs-based LFIA was successfully applied to food sample analysis, including milk and orange juice, demonstrating its robust applicability for real-world detection scenarios. This approach not only lowered the detection limit but also reduced the total detection time, making it a highly promising candidate for on-site detection of <i>Staphylococcus aureus</i> and other foodborne pathogens by simply changing the recognition element.</p> Graphical Abstract <p></p>

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Ultrasmall high-entropy alloy-nanolabels based immunochromatographic test strip for rapid, ultrasensitive, and catalytic detection of Staphylococcus aureus

  • Wei Huang,
  • Zhen Ren,
  • Xukui Li,
  • Rong Chen,
  • Dinglin Fan,
  • Jingjing Da,
  • Yan Zha,
  • Yongjie Xu

摘要

The use of ultrasmall high-entropy alloys (HEAs, ≈5 nm) with unique catalytic properties, integrating them into lateral flow immunoassay (LFIA) for the detection of Staphylococcus aureus is proposed. Under optimal conditions, the detection limits of the HEAs-based LFIA were 1.5 × 103 CFU/mL via direct coloration and 15 CFU/mL using the DAB-enhanced catalytic signal, representing improvements of 103-fold and 105-fold, respectively. The HEAs demonstrated superior performance compared with traditional AuNPs, including enhanced sensitivity and high peroxidase activity (Km = 0.066 M, Vmax = 1.132 × 10–6 M min−1), as well as excellent specificity and repeatability, with relative standard deviation (RSD) of less than 10%. Moreover, the incubation time for HEAs functionalized with antibodies to bind Staphylococcus aureus was only 5 min, compared with 20 min for functionalized AuNPs. The method also showed high specificity, effectively discriminating Staphylococcus aureus from other foodborne pathogens. The HEAs-based LFIA was successfully applied to food sample analysis, including milk and orange juice, demonstrating its robust applicability for real-world detection scenarios. This approach not only lowered the detection limit but also reduced the total detection time, making it a highly promising candidate for on-site detection of Staphylococcus aureus and other foodborne pathogens by simply changing the recognition element.

Graphical Abstract