Development of a Time-Resolved Fluorescent Microsphere Test Strip for Rapid, On-Site, and Sensitive Detection of...

Picoxystrobin (PIC) is a fungicide extensively used for disease control in both crops and vegetables. Residues of PIC in vegetables pose a potential threat to human health due to their accumulation in the food chain. In this study, a specific PIC monoclonal antibody (mAb) was developed by introducing a carboxylic acid arm into PIC and subsequently preparing a hapten and an artificial antigen. A sensitive and rapid time-resolved fluorescence immunochromatographic assay (TRFICA) was established based on the mAb. Subsequently, using a time-resolved fluorescent microsphere (TRFM) as signal probe, mAbs and microspheres were covalently coupled. The activated pH, the mAb diluents, the mAb amount, and the probe amount were optimized. Under optimized conditions, the quantitative limits of detection (qLOD) of PIC in cucumber, green pepper, and tomato using TRFICA were established at 0.61, 0.26, and 3.44 ng/mL, respectively; the 50% inhibiting concentrations (IC₅₀) were 11.76, 5.29, and 37.68 ng/mL, respectively. The linear ranges were 1.81-76.71, 0.80-35.04, and 8.32-170.55 ng/mL, respectively. The average recovery in cucumber, green pepper, and tomato samples ranged from 79.8% to 105.0%, and the corresponding coefficients of variation (CV) were below 14.2%. In addition, 15 vegetable samples were selected and compared with the results obtained using ultra-performance liquid chromatography/tandem mass spectrometry (UPLC-MS/MS). The results revealed a high degree of concordance between the proposed method and UPLC-MS/MS. In conclusion, the devised TRFICA method is a valuable tool for rapid, on-site, and highly sensitive detection of PIC residues in vegetables.