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.