Development of\nHighly Sensitive Immunochromatography\nUsing Time-Resolved Fluorescence Microspheres for Amantadine Detection

Amantadine (AMA), commonly used to treat viral infections\nin livestock\nand poultry, has been banned owing to its potential hazards to human\nwell-being. To detect unauthorized AMA usage in livestock, we developed\na polyclonal antibody with a high affinity for the specific recognition\nof AMA through a rational design based on a structure similar to AMA\nand revealed the availability of the hapten design by computational\nchemistry analysis. Using this antibody, we established a highly responsive\ntime-resolved fluorescence immunochromatographic assay (TRFICA). The\nvisual detection limit of the assay is 0.6 μg/kg, and the quantitative\ndetection limit is 0.05 μg/kg. The TRFICA also showed good recovery\nrates ranging from 94.5 to 109.9%, with variability coefficients not\nexceeding 10%. The outcomes of undisclosed sample examinations aligned\nwith those of HPLC-MS/MS analyses, indicating that this approach can\nfunction as an ideal screening and monitoring tool for detecting illegal\nAMA in chicken muscle.