Study on the Detection Method of Cloxacillin Visualized Immunoaffinity Gel Column
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Abstract:
In order to establish a method for the detection of cloxacillin visualized immunoaffinity gel column, the activated ester method was used to combine horseradish peroxidase (HRP) with cloxacillin to obtain the cloxacillin enzyme standard antigen. Results of the UV spectrum scanning showed that the CLOX enzyme standard antigen was successfully synthesized. The agarose gel activated by hydrogen bromide was combined with cloxacillin antibody and HRP antibody, respectively, which was used as the detection layer and the quality control layer of the affinity gel column, respectively. A visual immunoaffinity gel column analysis method for cloxacillin was established. The optimum condition of this method was the dilution ratio of enzyme-labeled antigen of 4000, dilution ratio of antibody gel of 10, dilution ratio of HRP gel of 20, sample dilution pH of 5.7 and sample addition time of 60 s. The detection limit of this method was 25 μg/L. Its specificity was strong. There was no cross-reaction with piperacillin, amoxicillin, ampicillin, tetracycline, furantanone, thiamphenicol and streptomycin. The detection limit of cloxacillin in animal-derived foods was 25 μg/kg. This method is convenient, rapid and visual. It is suitable for high-throughput screening of cloxacillin.
