Development of a High-Throughput Indirect Competitive Chemiluminescence Enzyme-Linked Immunoassay for the Rapid Detection of Bongkrekic Acid in Tremella Fungus and Rice Noodles

. This toxin is commonly found in spoiled fermented rice- and wheat-based products, snow fungus, and black fungus and can cause severe foodborne illness. The development of a rapid onsite detection method can effectively prevent food poisoning incidents and ensure food safety. In this study, a highly specific anti-BKA monoclonal antibody was prepared, the reaction conditions were optimized, and an indirect competitive chemiluminescent enzyme-linked immunoassay (ic-CLEIA) system was developed for high-throughput screening of BKA in food. The results showed that the ic-CLEIA had good linearity in the range of 7.3-106.6 pg/mL, a limit of detection of 4.7 pg/mL, a limit of quantification of 7.3 pg/mL, a half-maximal inhibition concentration of 28.2 pg/mL, a spike recovery of 86.6-94.1%, a coefficient of variation of less than 10%, and no cross-reactivity with structural analogs. There was no significant difference between the detection results obtained with ic-CLEIA and ultraperformance liquid chromatography-tandem mass spectrometry for the samples. This method provides reliable technical support for food safety monitoring, especially for grassroots laboratories and large-scale sample screening.