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United States Department of Agriculture

Agricultural Research Service

Title: Solid-Phase Immunosensor for Quantitative Rapid Detection of Antibiotic Residues in Raw Unprocessed Milk.

Authors
item Anis, Nabil - CVM, ROCKVILLE, MD
item Gotthardt, Joan - CVM, ROCKVILLE, MD
item Thomas, Michael - CVM, OR, LAUREL, MD
item Von Bredow, Jurgen - CVM, OR, LAUREL, MD
item Khalil, Maha - UNIV.OF MARYLAND,BALTIMOR
item Stanker, Larry
item Menking, Darrel - U.S. ARMY, ABERDEEN, MD
item Valdes, James - U.S. ARMY, ABERDEEN, MD
item Park, Jim - U.S. ARMY, ABERDEEN, MD

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: May 18, 2004
Publication Date: May 18, 2004
Citation: Anis, N., Gotthardt, J., Thomas, M., Von Bredow, J., Khalil, M., Stanker, L.H., Menking, D., Valdes, J., Park, J. 2004. Solid-phase immunosensor for quantitative rapid detection of antibiotic residues in raw unprocessed milk.. Meeting Abstract.

Technical Abstract: Solid-Phase immunosensor for quantitative rapid detection of antibiotic residues in raw unprocessed milk. Solid-phase immunosensor for quantitative rapid detection of antibiotic residues in raw unprocessed milk Contamination of milk by drug residues represents a major public heath hazard. Current methods for residue detection are either very slow (microbial growth assay) or laborious and time consuming (e.g., HPLC, GC, or GC-MS). A simple detection system that is cost effective and user-friendly is needed for rapid screening of milk samples for drug residues. A solid-phase competitive fluoroimmunoassays using a 12 channel fully automated flow fluorometer adopted for rapid screening of ceftiofur in raw unprocessed milk was developed. The fluorescent signal was generated by binding of fluorescein-labeled secondary antibodies (anti-mouse IgG-FITC) to a protein complex consisted of BSA-conjugated ceftiofur and monoclonal anti-ceftiofur antibodies immobilized on the surface of polymethyl-methacrylate (PMMA) beads. Binding of the anti-ceftiofur antibodies to the BSA-ceftiofur-coated beads was inhibited by the free ceftiofur in milk samples in a dose dependent manner. The binding of the mAb anti-ceftiofur to the BSA-ceftiofur-coated beads was specific as the mAb anti-ceftiofur was not bound to beads coated with non-specific BSA-antigen or beads coated with non-specific proteins such as casein, BSA, or IgG. The Limit of Quantitation (LOQ) of the assay was 0.3 ppb. The dynamic range of the assay was 1 ppb to 50 ppb. The assay discriminated effectively between ceftiofur at 5 ppb and other commonly used antibiotics which were not detected at 100 ppb. The assay provided direct, simple, rapid, and specific screening of milk samples without sample preparation that is usually needed for residue detection by instrumental analysis.

Last Modified: 10/26/2014
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