J Vet Sci.  2006 Jun;7(2):111-117. 10.4142/jvs.2006.7.2.111.

Development of immunoassays for the detection of kanamycin in veterinary fields

Affiliations
  • 1Institute for Zoonotic Disease, College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea.
  • 2Department of Biochemistry, College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea. vetlee@snu.ac.kr

Abstract

Monoclonal antibody against kanamycin was prepared, and competitive direct ELISA and immunochromatographic assay were developed using the antibody to detect kanamycin in animal plasma and milk. The monoclonal antibody produced was identified to be IgG1, which has a kappa light chain. No cross-reactivity of the antibody was detected with other aminoglycosides, indicating that the monoclonal antibody was highly specific for kanamycin. Based on competitive direct ELISA, the detection limits of kanamycin were determined to be 1.1 ng/ml in PBS, 1.4 ng/ml in plasma, and 1.0 ng/ml in milk. The concentration of intramuscularly injected kanamycin was successfully monitored in rabbit plasma with competitive direct ELISA. Based on the colloidal gold-based immunochromatographic assay, the detection limits of kanamycin were estimated to be about 6-8 ng/ml in PBS, plasma, and milk. The immunochromatographic assay would be suitable for rapid and simple screening of kanamycin residues in veterinary medicine. Screened positives can be confirmed using a more sensitive laboratory method such as competitive direct ELISA. Therefore, the assays developed in this study could be used to complement each other as well as other laboratory findings. Moreover, instead of slaughtering the animals to obtain test samples, these methods could be applied to determine kanamycin concentration in the plasma of live animals.

Keyword

competitive direct ELISA; immunochromatographic assay; kanamycin; monoclonal antibody

MeSH Terms

Animals
Anti-Bacterial Agents/*analysis
Antibodies, Monoclonal
Chromatography/methods/veterinary
Enzyme-Linked Immunosorbent Assay/methods/*veterinary
Kanamycin/*analysis
Mice
Milk/*chemistry
Rabbits
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