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Anti-idiotypic nanobody-phage display-mediated real-time immuno-PCR for sensitive, simultaneous and quantitative detection of total aflatoxins and zearalenone in grains
- Ren, Xianfeng, Zhang, Qi, Wu, Wenhua, Yan, Tingting, Tang, Xiaoqian, Zhang, Wen, Yu, Li, Li, Peiwu
- Food chemistry 2019 v.297 pp. 124912
- DNA, aflatoxins, bacteriophages, chemical species, detection limit, grains, high performance liquid chromatography, monoclonal antibodies, nucleotide sequences, polymerase chain reaction, zearalenone
- An anti-idiotypic nanobody-phage display-mediated immuno-polymerase chain reaction (PD-IPCR) method was developed for simultaneous quantitative detection of total aflatoxins and zearalenone in cereals. Two phages, displaying the variable domain of the heavy chain anti-idiotypic nanobody that binds aflatoxin- or zearalenone-specific monoclonal antibody (1C11 or 2D3), were used as competitors for corresponding analytes. Specific DNA sequences encoding anti-idiotypic nanobodies were used to design the primers for PCR amplification. The results indicated that detection limits for total aflatoxins and zearalenone in a sample were 0.03 and 0.09 ng mL−1, respectively. Recoveries of spiked aflatoxins and zearalenone were 80–118% and 76.7–111%, respectively. Validation results were in good agreement with the gold-standard high-performance liquid chromatography method. This report is the first to describe PD-IPCR for simultaneous quantitative detection of total aflatoxins and zearalenone in cereals.