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The zearalenone (ZEA) monoclonal antibody (mAb) 2D3, one of the highest sensitivity antibodies, was developed. Based on this mAb, it was established of an immunoaffinity column (IAC) coupled with an indirect competitive enzyme-linked immunosorbent assay (icELISA). After optimization, the icELISA allowed an IC50 against ZEA of 0.02 µg L−1. The mAb 2D3 exhibited a high recognition of ZEA (100%) and β-zearalenol (β-ZOL, 88.2%). Its cross-reactivity with α-zearalenol (α-ZOL) and β-zearalanol (β-ZAL) were found to be 4.4% and 4.6%, respectively. The IAC-icELISA method was employed to analyze ZEA contamination in food samples, compared with high-performance liquid chromatography (HPLC). The spiked assay for ZEA demonstrated the considerable recoveries for IAC-icELISA (83–93%) and HPLC (94–108%) methods. Results showed that the mAb 2D3 and IAC-icELISA method posed potential applications in sensitively determination of ZEA in maize. [ABSTRACT FROM AUTHOR]

To simultaneously detect two metabolites of Aspergillus flavus, namely, cyclopiazonic acid (CPA) and aflatoxin (AFT), an ultrasensitive monoclonal antibody (mAb) YTT-2 against CPA was developed and characterized, with sensitivity to CPA of 1.32 ng mL−1. Along with the previously homemade mAb 1C11 against AFT, two mAbs were used to develop time-resolved fluorescence immunoprobes or gold immunoprobes. We developed two multiple-analyte paper immunosensors including time-resolved fluorescent immunochromatographic assay (TRFICA) and gold immunochromatographic assay (GICA) for the simultaneous determination of CPA and AFT. The TRFICA showed limits of determination (LODs) of 0.21 and 0.004 ng mL−1, while the GICA showed LODs of 0.33 and 0.01 ng mL−1 for CPA and AFT, respectively. To validate the specificity of the two rapid immunoassays, rice, corn and peanut samples were spiked with different concentrations of CPA and AFT. The two methods showed satisfactory recoveries (76.39~90.82% for CPA and 84.60~94.45% for AFT) and coefficients of variation of 3.50~7.80% for CPA and 4.12~13.90% for AFT. The results indicated that the TRFICA could complete the test within 5 min and had lower LODs and linear ranges, compared with that of GICA. The method developed in this work can be widely applied to the rapid and quantitative simultaneous determination of multiple harmful metabolites in fungi for food safety and health care. [ABSTRACT FROM AUTHOR]

Abstract: Mycotoxins are highly toxic contaminants and have induced health threat to human and animals. Aflatoxin B1 (AFB1), ochratoxin A (OTA) and zearalenone (ZEA) commonly occur in food and feed. A multi-component immunochromatographic assay (ICA) was developed for rapid and simultaneous determination of these three mycotoxins in agro-food. The strategy was performed based on the competitive immunoreactions between antibody-colloidal gold nanoparticle conjugate probes and mycotoxins or mycotixin antigens. Each monoclonal antibody specially recognize its corresponding mycotoxin and antigen, and there was no cross reactivity in the assay. Three mycotixin antigens were immobilized as three test lines in the nitrocellulose membrane reaction zone, which enable the simultaneous detection in one single test. The visible ICA results were obtained in 20min. The visual detection limits of this strip test for the AFB1, OTA and ZEA were 0.25ng/mL, 0.5ng/mL and 1ng/mL, respectively. The assay was evaluated using spiked and naturally contaminated peanuts, maize and rice samples. The results were in accordance with those obtained using enzyme-linked immunosorbent assay. In summary, this developed ICA could provide an effective and rapid approach for onsite detection of multi-mycotoxin in agro-food samples without any expensive instrument. [Copyright &y& Elsevier]