A Highly Sensitive and Specific SERS-Based Lateral Flow Immunochromatographic Assay Using Encoded Silver-Coated Petal-Like Gap-Enhanced Raman Tags (P-GERT@Ag) for Simultaneous Detection of T-2 Toxin and Deoxynivalenol in Food Samples on a Single Test Line

T-2 toxin and deoxynivalenol (DON) are major trichothecene mycotoxins that frequently contaminate agricultural products and pose substantial risks to human and animal health. Here, a highly sensitive and specific surface-enhanced Raman scattering lateral flow immunochromatographic assay (SERS-LFIA) was developed for simultaneous determination of T-2 toxin and DON on a single test line. Silver-coated petal-like gap-enhanced Raman tags encoded with 4-aminothiophenol or 4-nitrobenzenethiol were conjugated with antibodies against T-2 toxin or DON, respectively. A mixture of T-2 toxin-ovalbumin and DON-ovalbumin coating antigens was immobilized on the test line. After a 15 min assay, Raman signals at 1136 cm−1 and 1333 cm−1 were measured for quantitative detection. The assay produced IC50 values of 0.35 ng mL−1 for T-2 toxin and 0.29 ng mL−1 for DON, with limits of detection of 0.0033 and 0.0067 pg mL−1, respectively. No mutual interference or significant cross-reactivity with aflatoxin B1, ochratoxin A, zearalenone, patulin, or fumonisin B1 was observed. Recoveries from spiked corn and wheat samples ranged from 89.7% to 114.6%, with relative standard deviations below 9.09%. This rapid platform therefore enables sensitive and specific multiplex mycotoxin analysis on a single test line.