Broad-specificity antibody profiled by hapten prediction for immunoassay development of anilinopyrimidine fungicides

Improper application of anilinopyrimidine (AP) fungicides on fruits and vegetables may lead to residues that threaten human health through the food chain. Developing rapid detection methods for AP fungicides is therefore critical for food safety supervision. Herein, a computer-aided hapten design strategy was employed to design broad-spectrum haptens against cyprodinil (Cyp), mepanipyrim (Mep), and pyrimethanil (Pyr), enabling the generation of broad-spectrum anti-AP monoclonal antibodies. An immunochromatographic assay (ICA) was further developed for simultaneous rapid detection of Cyp, Mep, and Pyr residues in grapes, strawberries, and tomatoes, with calculated limits of detection of 0.02-0.07 μg/kg. The ICA exhibited sufficient sensitivity for routine detection, and matrix spike recoveries ranged from 84.7%-94.7%. GC-MS/MS and authentic sample analysis confirmed its high accuracy and reliability, making it suitable for on-site screening.