Simultaneous and rapid detection of six different mycotoxins using an immunochip

Mycotoxins are highly toxic contaminants in food, animal feed, and commodities. The study has developed an immunochip for quantifying the concentrations of six mycotoxins: aflatoxin B1, aflatoxin M1, deoxynivalenol, ochratoxin A, T-2 toxin, and zearalenone, which were added to drinking water. The complete antigens (Ags) of the mycotoxins were contact printed and immobilized onto agarose-modified glass slides with 12 physically isolated subarrays, based on the reaction of both diffusion and covalent bond. The optimal concentration of each antigen and antibody (Ab) was obtained using an Ag–Ab immunoassay. Based on the indirect competitive immunoassay for the simultaneous detection of six mycotoxins in one single chip, six standard curves with good logistic correlation (R2 > 0.97) were respectively plotted. The working ranges (0.04–1.69, 0.45–3.90, 20.20–69.23, 35.68–363.18, 0.11–1.81, and 0.08–7.47 ng/mL, respectively) were calculated, as well as the median inhibitory concentrations (0.31 ± 0.04, 1.49 ± 0.21, 34.54 ± 1.30, 134.06 ± 11.75, 0.49 ± 0.05, and 1.54 ± 0.22 ng/mL, respectively), when six mycotoxins were detected simultaneously. Finally, the recovery rates in drinking water generally ranged from 80% to 120% on the same chip, with an intra-assay coefficient of variation lower than 15%. We successfully established an immunochip for simultaneous detection of six mycotoxins within 4 h, with advantages of using minimal samples and being visually semiquantitative with our naked eyes. In summary, the method could be developed on one single chip for detecting multiple contaminants in actual samples.

► We established an immunochip for detecting multiple targets simultaneously and rapidly.
► The immunochip was applied in drinking water with six different mycotoxins spiked.
► It is visual and semiquantitatve for the immunochip to detect six mycotoxins.
► Due to diffusion and covalent couple, probes with good activity can be immobilized onto agarose-modified chips.
► Their detection limits are at the level of picogram or nanogram.