| Fluoroquinolones(FQNs) medicine is a kind of artificial synthetic neotype antibacterials,along with its general use in healing infectious diseases of mankind and animals, the residue problem has caused much attention of people. At present, commonly used detective methods are HPLC,ELISA, et al. Among the total, ELISA is easy to operate and has high sensitivity, refer to detect residual medicine generously and quickly. This research intend to establish a method of the indirect competive ELISA to detect residue.This experiment firstly coupled ENRO,NFLX,DFLX, which are three commonly used FNQs medicine, with BSA and OVA by NHS, formed complete antigen differently, then identified the complete antigen by ultraviolet scan spectrographical identification.Immuned New Zealand albino rabbit to prepare respective antiserum. Then operated competive reaction with other commonly used medicine(Enrofloxacin,Sarafloxacin,Norfloxacin,Danofloxacin,Ciprofloxacin,Lomefloxacin,Ofloxacin,Peflacine)respectively through indirect competive ELISA. Calculated consensual reaction rate, and bolting DFLX, which could synchronize contest reaction with itself and other five kinds of FQNs medicine: SALX,ENRO,NFLX,LMLX,OFLX simultaneously,and the consensual reaction rate was high.After optimized the condition of ELISA, the optimal concentration of the coating antigen were 1.25μg/L, the working concentration of DFLX-PcAb and sheep anti- rabbit IgG were 1:10000 and 1:4000 respectively, then got the regression equation (y=0.7975-0.125x R2=0.989) and the most suitable detect range(0.1～10 ng /L). Through evaluated this method, gained the coefficients of variation of intra-assay and inter-assay were 3.81% and 6.25%; degree of accuracy (standard recovery rate)was 104.41%; sensibility(lowest detectable limit)was 0.8ng/mL. All the index accorded with the demand of indirect competitive ELISA, so the indirect competitive ELISA was established to detect the residues of DFLX.
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