Development of ELISA and immunochromatographic assay for the detection of neomycin.
BACKGROUND
Reliable analytical methods are required to monitor neomycin residue levels in the livestock products. In particular, a more simple and rapid detection method is required in the veterinary fields.


METHODS
Competitive direct ELISA and immunochromatographic assay were developed using monoclonal antibody to detect neomycin in the animal plasma and milk.


RESULTS
No cross-reactivity of the antibody was observed with other aminoglycosides based on competitive direct ELISA methods, indicating that the antibody is highly specific for neomycin. Based on the standard curves, the detection limits were determined to be 6.85 ng/ml in PBS, 3.61 ng/ml in plasma, and 2.73 ng/ml in milk, respectively. Recoveries of neomycin from spiked plasma and milk at levels of 50-200 ng/ml ranged from 87% to 108%. Concentration of intramuscularly injected neomycin was successfully monitored in the rabbit plasma through competitive direct ELISA. Immunochromatographic method was also developed using colloidal gold-conjugated monoclonal antibody. Through this method, the detection limits were estimated to be about 10 ng/ml of neomycin in PBS, plasma, and milk.


CONCLUSIONS
Immunochromatographic assay developed in this study is suitable for the simple screening of neomycin residues in the veterinary field. Observed positives can be confirmed using a more sensitive laboratory method such as competitive direct ELISA.
Jin
Yong
Y
Department of Biochemistry, College of Veterinary Medicine, Seoul National University, 151-742 Seoul, South Korea.
Jang
Jin-Wook
JW
Lee
Mun-Han
MH
Han
Chang-Hoon
CH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D002845:Chromatography / Q000295:instrumentation / Q000379:methods* / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000662:veterinary; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D008892:Milk / Q000737:chemistry; D009355:Neomycin / Q000032:analysis* / Q000097:blood / Q000276:immunology; D011817:Rabbits; D015203:Reproducibility of Results
D000911:Antibodies, Monoclonal; D009355:Neomycin