Highly Sensitive Simultaneous Detection of Multiple Mycotoxins Using a Protein Microarray on a TiO2-Modified Porous Silicon Surface

A new protein microarray method for multiplex mycotoxin detection in parallel has been established on a stable TiO2-modified porous silicon (PSi) surface. A typical competitive immunoassay microarray protocol has been developed for simultaneous detection of multiplex mycotoxins including aflatoxin B1 (AFB1), ochratoxin A (OTA), and fumonisin B1 (FB1) on the TiO2-PSi surface. The epoxy groups were selected to modify the surface of a TiO2-PSi wafer for the immobilization of artificial antigens of mycotoxins because of their high signal-to-noise ratios. Under optimal conditions, the developed method showed wide linear detection ranges of 0.01–1 ng/mL for OTA, 0.001–1 ng/mL for AFB1, and 0.01–1 ng/mL for FB1 and low limit of detections (LODs) of 0.433 ng/mL for OTA, 0.243 ng/mL for AFB1, and 0.093 ng/mL for FB1. The microarray method can specifically identify the three mycotoxins and their analogues. The recovery rates in real samples were within 75–120%, which were in agreement with that of the classical ELISA method. The new method has great application potential for rapid, sensitive, and high-throughput screening of multiplex mycotoxins and other target molecules.