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

A new protein microarray method for multiplex mycotoxin detection in parallel has been established on a stable TiO-modified porous silicon (PSi) surface. A typical competitive immunoassay microarray protocol has been developed for simultaneous detection of multiplex mycotoxins including aflatoxin B (AFB), ochratoxin A (OTA), and fumonisin B (FB) on the TiO-PSi surface. The epoxy groups were selected to modify the surface of a TiO-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 AFB, and 0.01-1 ng/mL for FB and low limit of detections (LODs) of 0.433 ng/mL for OTA, 0.243 ng/mL for AFB, and 0.093 ng/mL for FB. 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.