Research On The Immunochromatographic Test Strip Of Flumequine

In this study,we choosed Flumequine(FLU)as the target,prepared capable of specifically recognizing Flumequine polyclonal antibody.The colloidal gold and colloidal carbon labled immunochromatographic test strip were established,respectively.Flumequine strips were applied in visual detecting flumequine residual in animal food.At last,a multi-residue strip with colloidal gold labeled salbutamol and colloidal carbon labeled flumequine was developed.Carbodiimide method and mixed anhydride method were prepared for immunogen FLU-BSA and FLU-KLH,immunizing New Zealand white rabbits obtained anti-flumequine polyclonal antibodies.Comparing titer and inhibition,the antibody immunized by immunogen FLU-BSA and produced by 1st rabbits was selected.The antiserum titer was 1:256000.An indirect competitive ELISA was developed.Under optimum operating conditions and the sensitivity IC50 and the detection limit IC15 were 10.9± 2.62 ?g/L and 0.79 ± 0.33 ?g/L,respectively.An immunochromatographic test strip of flumequine was established labeled with colloidal gold.The particles size of colloidal gold was 20 nm;the optimum pH gold-labeled was 8.5,the amount of antibody was 20 ?g/mL;the concentration of capture antibody was 0.5 mg/mL,the secondary antibody was diluted to 1:200,choosed Millipore HF135s models of nitrocellulose membrane as solid phase carrier,gold-labeled antibody was diluted 1:10 sprayed gold pad.The detection limit was 20 ?g/L.The seven kinds of real samples chicken,beef,pork,fish,shrimp,liver and milk were chosen to the spiked experiment,it was found that the detection limit was 50 ?g/L in sample.Secondly,immunochromatographic strip of flumequine was developed labeled with colloid carbon.Under optimal conditions,the carbon-labeled antibody spaying on conjugate release pad was infeasible;the detection limit was 20 ?g/L by carbon-labeled antibody mixed with the standard sample.The seven kinds of real samples chicken,beef,pork,fish,shrimp,liver and milk spiked experiment,the detection limit was 50 ?g/L in sample.A multi-residue strip was assembled with colloidal gold labeled salbutamol and colloidal carbon labeled flumequine.After the optimization,the results was clear and visual,red stripe representing salbutamol,the detection limit was 2 ?g/L;black stripe representing flumequine,the detection limit was 20 ?g/L.