Abstract:

Indirect competitive ELISA was adopted to develop a rapid and specific assay capable of detecting kanamycin. The optimal working conditions were as follows: coating buffer, 0.01 mol/L PBS solution at pH 7.4; confining buffer, 0.05 mol/L carbonate-bicarbonate buffer containing 0.1% casein solution; antibody dilution buffer, PBST buffer containing 4% polyethylene glycol 6000 at pH 8.5; dilution of secondary antibody, 1:3000; and standard dilution buffer, the same to coating buffer. The IC50 of kanamycin was 9.9 ng/ml, the detection limit 1.0 ng/ml, and the linear detection range 1－200 ng/ml.

Key words: kanamycin, ELISA, antibody, direct competition, indirect competition