Incorrect use of
tylosin and
tilmicosin could result in
allergy and select resistance. To monitor the illegal use of these
antibiotics in animals, a monoclonal-based indirect competitive
enzyme-linked
immunosorbent assay (ic-ELISA) has been established. Several
haptens were synthesized and conjugated to
carrier protein. Female Balb/c mice were inoculated with the four different conjugates to produce
monoclonal antibodies according to the schemes of immunization. Aftercell fusion and culture several times, nine hybridoma cell lines were isolated. Only one, 3C4 that has isotype
IgG2a, was selected for detailed study. The cross-reactivity of the
monoclonal antibody 3C4 to
tylosin and
tilmicosin was 100% and 51% respectively. The standard curves based on the
tylosin and
tilmicosin matrix calibration ranged from 2.5 to 40 μg L(-1), with an IC(50) value of 6.1 μg L(-1) and 12.1 μg L(-1), respectively. The limits of detection of the ic-ELISA ranged from 5.1 μg kg(-1) to 13.8 μg kg(-1) in edible animal tissues. The recoveries were 74.1% to 120.7% with less than 18.6% of the coefficient of variation when
tylosin and
tilmicosin were spiked in various biological matrices with the concentrations of 25.0-200.0 μg kg(-1). Good correlations between the results of the ic-ELISA and high performance liquid chromatography were observed in the incurred tissues. These results suggest that the ic-ELISA is a sensitive, accurate and low-cost method that would be a useful tool for the screening of the residues of
tylosin and
tilmicosin in muscle, liver, milk, honey and eggs.