| Objectives:To develop a competitive inhibition enzyme-linked immunosorbent assay (ciELISA) and colloidal gold probe-based immunochromatographic assay for the detection of AMOZ in tissue sample by the the monoclonal antibodies (mAbs) against the Metab olite of Furaltadone,5-morpholino-3-amino-2-oxazolidone (AMOZ).Methods:Firstly, AMOZ was reacted with 4-CBA and the hapten CPAMOZ was sy nthesized.The derivates were conjugated with BSA and OVA for complete antigens by using activated ester method. SDS-PAGE and the ult-raviolet spectra were used to identif y the artificial antigens. Five BALB/c mices were inmunized with CPAMOZ-BSA and th en the hybridom cell lines secreting monoclonal antibody against CPAMOZ were establis hed by using cell fusion, ELISA screening and subcloning. The immunological traits of t he mAbs such as the titer, subtype and the specificity were characterized in the way of indirect enzym-elinked immunosorbent assay (iELISA) and ciELISA.The ciELISA and col loidal gold immunochromatographic assay were established for AMOZ detection in tissue sample.Results:1.The competitive and complete antigens of AMOZ were synthesized succes sfully.2. After immunization and cell fusion, one hybridoma which can stably secrete m Abs against CPAMOZ was obtained.3. The secrete antibody was IgG1, the antibody tite r was up to 1×105, had less cross reactivity with other nitrofurans metabolites and deriva tes showing high specificity to CPAMOZ.3.The ciELISA for AMOZ detection was devel oed with 0.86μg/L of IC50 value to CPAMOZ and 0.07μg/L of LOD, the recovery of tissue samples was 81.48%-85.24%.The LOD of colloidal gold probe-based immunochrom atographic assay is 50μg/L.Conclusion:1.The complete antigens of AMOZ and the monoclonal antibody against CPAMOZ with high sensitivity and specificity had been generated successfully.2.The ci ELISA and colloidal gold immunochromatographic assay for AMOZ detection in tissue s ample were established successfully.
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