Development And Optimization Of Indirect Competitive ELISA Products Of Aflatoxin B₁

Aflatoxin B1 was found to be the most toxic and carcinogenic chemical substances, it is mainly caused by the metabolites of Aspergillus flavus and Aspergillus parasiticus together. Physical and chemical properties of aflatoxin B1 is stable and easy to contamination of agricultural products, food and feed. Aflatoxin invasion into the food chain, will produce the corresponding toxin. Human or animal consumption will have a great harm to the body. Currently, the world of agricultural products, food, feed and other items in the content of aflatoxin B1 toxin has strict limits. Avoid aflatoxin B1 contamination in the food chain, not only to prevent and control in agricultural raw materials, but also in the agricultural product material storage, food processing, logistics and distribution links to detect and monitor.In our country, the detection technology of aflatoxin B1 has been studied for many years, and has developed 3 kinds of different detection techniques. The international authoritative testing technology is the high performance liquid chromatography method, and our country also approved for this detection method, but this method is high cost, is not conducive to the operation, cumbersome pre-treatment of the sample, the overall testing process long time, is not conducive to monitoring department of the market dynamic supervision. Indirect competitive ELISA method is an experimental technique that uses an antigen and antibody interactions, enzyme-labeled secondary antibody was added with the substrate color reactions to determine the content of the test object. The experimental technique is designed to be a kit will be the promotion of products, it has to sample pre-treatment requirements low, easy to operate, and can greatly shorten the detection time advantage.The project developed the indirect competitive ELISA kit, necessary for optimum coating antigen added at 50ng/hole, aflatoxin B1 monoclonal antibody was added at 100ng/hole, the most appropriate secondary antibody dilution of 1:500. Its test results with the HPLC comparison coincidence rate greater than 98%, which proved consistent with the internationally accepted method of detection results. IC-ELISA kit to study accuracy, it took the detection of rice, corn, peanut cake toxin aflatoxin B1 detected, the detection result of the recovery of rice appear as:85%-91%, the recovery rate is displayed as corn detection:84% -94%, peanut cake detect recoveries read:88% -92%. Their recoveries are in line with national standards recoveries of 70%-120%, which illustrates the accuracy of the test results kits. The kit has been studied for sensitivity, detection limit his analysis of rice, corn, peanut cake. The results showed that the detection limit of rice show:0.25 μg/kg, corn detection limit display:0.30 μg/kg, the limit of detection peanut cake display:0.30 μg/kg, which illustrate the kit detection limits than the state standard lower limit of detection is more sensitive. For the study kit precision, taking a different ELISA assay plates aflatoxin B1 content detection of aflatoxin in rice, which showed differences within the panel 3.0%、2.7%、3.2%, the difference between the plates is 2.9%. It describes the development of the same batch of kits and kits of different batches of test results is insignificant, its detection method is stable, accurate and effective.