Rapid Detection of Deoxynivalenol in Cereals Based on Amplified Luminescence Proximity Homogeneous Detection Technology

Background: To address challenges of low sensitivity, complex operation, and matrix interference in detecting deoxynivalenol (DON) in cereals, this study developed a novel wash-free detection system using amplified luminescence proximity homogeneous assay (AlphaLICA) technology. Methods: The method leverages energy transfer between microspheres and antigen-antibody interactions: DON antibodies competitively bind to DON-BSA-luminescent microspheres and DON standards/samples, with bound antibodies detected via goat anti-mouse IgG-conjugated photosensitive microspheres. Results: Results demonstrated a detection limit (LOD) of 1.03 ng/mL and quantitation limit (LOQ) of 7.57 ng/mL. Intra-/inter-batch coefficients of variation were <6.13% and 6.67%, respectively. Recovery rates in wheat, corn, and rice reached 97.52%–105.83% (relative standard deviation (RSD) ≤12.83%), with <0.21% cross-reactivity to aflatoxin B1. Compared to enzyme-linked immunosorbent assay (ELISA), AlphaLICA showed superior sensitivity (1.03 vs. 10 ng/mL), faster detection (5 vs. 35 min), broader linear range (7.57–800 vs. 12.5–200 ng/mL), and lower variability (6.13%/6.67% vs. 8%/12%). Pearson correlation analysis confirmed a strong association with ELISA results (p < 0.001, r = 0.8374). Conclusions: The established DON-AlphaLICA method provides a simple, rapid, sensitive, and accurate solution for DON detection, effectively addressing current limitations in grain safety monitoring. This innovative method enables efficient screening of mycotoxin contamination in agricultural products while minimizing procedural complexity.