Development of ELISA Tchnique for Fumonisin B1 Detection in Corn

Abstract

 Enzyme-Linked Immunosorbent Assay *ELISA) was developed for Fumonisin B1 detection.  Polyclonal antibodies against Fumonisin B1 (FmB1) were produced in teh rabbits after immunizing the animal with FmB1-brovine serum albumin.  The antibody against FmB1 was obtained from the rabbit after 5 weeks of immunization.  The highest antibody titer was found in the sera of rabbit at the 10th week with the titer of 1:25,000.  The preparation of FmB1-HRP conjugate was achieved by a water soluble carbodiimide method.  A direct competitive ELISA was developed using the produced anibody and and FmB1 -HRP conjugate at the concentration of 1:1,000 and 1:2,000 respectively for detection of the FmB1 in corn samples.  The appropriate time for teh reaction between FmB1 and FmB1 -HRP in competition to bind the antibody at the bottom of the well was 60 minutes.  High correlation of ELISa with HPLC data for detection of FmB1 in naturally contaminated corn samples was obtained with the linear regression equation of y = 0.8943x + 0.0904.  and correlation coefficient of 0.97.  The ELISA technique for Fumonisin B1 detection in corn was successfully developed in this studies and should be further developed as commercial ELISA test kit.