A Highly Sensitive and Group-Specific Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection of AFB 1 in Agriculture and Aquiculture Products.

Aflatoxins (AFs) including AFB 1 , AFB 2 , AFG 1 and AFG 2 are widely found in agriculture products, and AFB 1 is considered one of the most toxic and harmful mycotoxins. Herein, a highly sensitive (at the pg mL -1 level) and group-specific enzyme-linked immunosorbent assay (ELISA) for the detection of AFB 1 in agricultural and aquiculture products was developed. The AFB 1 derivative containing a carboxylic group was synthesized and covalently linked to bovine serum albumin (BSA). The AFB 1 -BSA conjugate was used as an immunogen to immunize mice. A high-quality monoclonal antibody (mAb) against AFB 1 was produced by hybridoma technology, and the mAb-based ELISA for AFB 1 was established. IC 50 and limit of detection (LOD) of the ELISA for AFB 1 were 90 pg mL -1 and 18 pg mL -1 , respectively. The cross-reactivities (CRs) of the assay with AFB 2 , AFG 1 , and AFG 2 were 23.6%, 42.5%, and 1.9%, respectively, revealing some degree of group specificity. Corn flour, wheat flour, and crab roe samples spiked with different contents of AFB 1 were subjected to ELISA procedures. The recoveries and relative standard deviation (RSD) of the ELISA for AFB 1 in spiked samples were 78.3-116.6% and 1.49-13.21% ( n = 3), respectively. Wheat flour samples spiked with the mixed AF (AFB 1 , AFB 2 , AFG 1 , AFG 2 ) standard solution were measured by ELISA and LC-MS/MS simultaneously. It was demonstrated that the proposed ELISA can be used as a screening method for evaluation of AFs (AFB 1 , AFB 2 , AFG 1 , AFG 2 ) in wheat flour samples.