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• A duplex phage-display immuno-polymerase chain reaction (PD-IPCR) was developed. • PD-IPCR was shown to be a sensitive method for mycotoxins detection. • The PD-IPCR was used successfully to evaluate aflatoxins and zearalenone in grains. 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. [ABSTRACT FROM AUTHOR]