Multiplex Flow Cytometric Immunoassays for High-Throughput Screening of Multiple Mycotoxin Residues in Milk

Microsphere-based immunoassays involving flow cytometry have recently gained popularity for use in protein detection and infectious disease diagnosis due to their simple assay format and capacity for multiplexed analysis. In this work, four mycotoxins, including aflatoxin M1 (AFM1), ochratoxin A (OTA), deoxynivalenol (DON), and fumonisin B1 (FB1), were selected as model analytes. A quadplex flow cytometric immunoassay (FCIA) was developed for detecting multiple mycotoxin residues in milk. The optimized quadplex FCIA exhibited satisfactory compatibility with the multiple mycotoxin residue analysis, with limits of detection (LODs) of 0.045 μg L−1 for AFM1, 0.94 μg L−1 for OTA, 7.48 μg L−1 for DON, and 2.45 μg L−1 for FB1. The recoveries of the target mycotoxins from spiked milk were 76–95%, with a relative standard deviation of less than 13.4%. Compared to traditional ELISA formats, the FCIA provided lower detection limits for multiple mycotoxin detection.