| Aflatoxin B1 is the secondary metabolites which was mainly produced by A spergillus flavus, Aspergillus parasiticus and Aspergillus nomius. It widely were found in peanuts, cottonseed, corn, wheat and rice and other crops. It is the most chemical carcinogens. It was extremely toxic and carcinogenic. It can caused animal liver carcinogenic, stomach carcinogenic and kidney carcinogenic, but also threated human and animal husbandry through the food chain. There were very strict standard limits of AFB1 in tvarious types of food in many countries.Therefore, it was important practical significance to study an accurate, sensitive and rapid method of detection AFB1. Many department have researched a lot detection methods for AFB1 in many countries.This study established a indirect competition chemiluminescence enzyme immunoassay (Chemiluminescence enzyme immunoassay, CLEIA) detection of AFB1 in food. The main contents were as follows:1 By analysis of the antigen coating concentration, closed conditions, antibody concentration and reaction time in competitive reaction system, the optimal reaction conditions were defined. Coating AFBi-BSA antigen concentration was 1.5μg/ml. Monoclonal anti-AFB1 antibody dilution was 1:150000. Optimal dilution of HRP secondary antibody degree was 1:2000. Competitive reaction time was 45min.2 The detection limit was 8.68 pg/ml. The linear range of calibration curve were 15.63 pg/ml-500.00 pg/ml. IC50 was 54.96 pg/ml, the linear regression equation: y=23.264x-43.206, R2=0.9924; The average intra-and inter-coefficients of variation were 4.03% and 6.97%. The average recovery was 92.72%. The average coefficient of variation was 8.68%.3 The 32 of samples were individual determined by indirect competition AFB1-CLEIA and purchased AFB1-ELISA quantitative determination test kits.The two methods showed no significant difference P<0.01, correlation coefficient R2= 0.9954. |
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