A Hierarchical Au@PDA-PdCu Nanozyme for Signal-Amplified Immunochromatographic Detection of Carbofuran in Leafy Vegetables

Abstract Carbofuran residues threaten food safety and public health, yet conventional instrumental methods require expensive equipment and complicated pretreatment, while traditional immunochromatographic assays still lack sufficient sensitivity. Herein, a hierarchical Au@PDA-PdCu nanozyme was developed as a signal probe for a competitive immunochromatographic assay (ICA) for rapid carbofuran detection. The Au core served as a stable nanoplatform, the PDA interlayer enabled antibody immobilization, and the PdCu shell provided strong peroxidase-like activity for post-assay signal amplification. Three probe formats, namely liquid, sprayed, and lyophilized probes, were constructed and optimized. After DAB enhancement, the detection limit of the liquid-format strip decreased from 0.055 to 0.016 ng/mL, while the linear ranges of the sprayed- and lyophilized-format strips expanded to 0–100 ng/mL. In Chinese cabbage, chive, and spinach, all three formats exhibited good quantitative performance (R² > 0.98). The visual limits of detection were 5 ng/mL for the liquid- and sprayed-format strips and 1 ng/mL for the lyophilized-format strip. Recoveries ranged from 70% to 119%, with coefficients of variation of 3.7%–25.7%. Good specificity was observed, with cross-reactivity below 0.1% for most tested compounds except 3-hydroxycarbofuran. The liquid and lyophilized strips showed predicted shelf lives of over 1 year, and ICA results agreed well with LC–MS/MS. Overall, this method provides a rapid, sensitive, and practical tool for on-site carbofuran screening in leafy vegetables.