Determination Of Penicillin Content In Milk By ELISA And Preperation Of ELISA Kits

Since the invention of penicillin, a large number of antibiotics were widely used due to their excellent antibacterial effect, low toxicity and side effects. But it is found in recent years that the transition misuse of antibiotics has led to many serious impacts on human health.Wherein the milk product in question for the prevention and treatment of mastitis cows led to excessive use of penicillin residues in excess of penicillin -related problems in dairy products, has attracted the attention States drug regulatory authorities, and dairy products allowed content of penicillin antibiotics a clearly defined.In order to to explore an effectiv, accurate and convenient method for the detection of penicillin content goals, a test enzyme-linked immunosorbent assay kit was developed in this study.The process for separation of penicillinase from Bacillus cereus was built. Firstly, an affinity chromatography medium was prepared by connecting the ampicillin to the gen bromide-activated Sepharose, and then the high activity penicillinase from Bacillus cereus was obtained by salting-out, affinity chromatography, dialysis and other complex process. Nextly, based on enzyme-linked immunoassay technology, the penicillinase coated on the microtiter plate was formed by polystyrene according to the principle of competition kit. Ampicillin with horseradish peroxidase is preferably prepared for the detection of active enzyme conjugate. Then optimize the solution formulation, developed to detect levels of penicillin in milk sensitivity to 4ng/ml (European standard) ELISA kit.