Development of ELISA and immunochromatographic assay for the detection of gentamicin.
Competitive direct enzyme-linked immunosorbent assay (ELISA) and the immunochromatographic assay were developed using a monoclonal antibody to detect gentamicin in the animal plasma and milk. No cross-reactivity of the antibody was observed with other aminoglycosides based on competitive direct ELISA, indicating that the antibody is highly specific for gentamicin. On the basis of the standard curves, the detection limits were determined to be 0.9 ng/mL in phosphate-buffered saline (PBS), 1.0 ng/mL in plasma, and 0.5 ng/mL in milk, respectively. Recoveries of gentamicin from spiked plasma and milk at levels of 25-100 ng/mL ranged from 85 to 112%. The concentration of intramuscularly injected gentamicin was successfully monitored in the rabbit plasma through competitive direct ELISA. The detection limits were estimated to be about 6 ng/mL of gentamicin in PBS, plasma, and milk using the colloidal gold-based immunochromatographic assay, which is suitable for the simple screening of gentamicin residues in the veterinary field. Observed positives can be confirmed using a more sensitive laboratory method such as competitive direct ELISA. Therefore, the assays developed in this study could complement each other as well as veterinary field and laboratory findings.
Jin
Yong
Y
Department of Biochemistry, College of Veterinary Medicine, Seoul National University, 151-742 Seoul, Republic of Korea.
Jang
Jin-Wook
JW
Han
Chang-Hoon
CH
Lee
Mun-Han
MH
D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000097:blood; D000911:Antibodies, Monoclonal; D002845:Chromatography / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D005839:Gentamicins / Q000032:analysis* / Q000097:blood; D007118:Immunoassay / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D008892:Milk / Q000737:chemistry
D000900:Anti-Bacterial Agents; D000911:Antibodies, Monoclonal; D005839:Gentamicins