Multiplex SERS-based lateral flow immunosensor for the detection of major mycotoxins in maize utilizing dual Raman labels and triple test lines.

A multiplex surface-enhanced Raman scattering (SERS)-based lateral flow immunosensor was developed to determine six major mycotoxins in maize. Two characteristic Raman reporter molecules-5,5-dithiobis-2-nitrobenzoic acid (DTNB) and 4-mercaptobenzoic acid (MBA)-were used to label the synthesized Au@Ag core-shell nanoparticles for the preparation of SERS nanoprobes as detection reagents. Six corresponding hapten-protein conjugates were prepared and dispensed on three test lines of nitrocellulose membrane with two conjugates on each line as capture antigens. This design facilitates the simultaneous detection of the six mycotoxins in a single test. After optimizing the experimental parameters of immunosensor, the limits of detection were as low as 0.96 pg/mL for aflatoxin B1, 6.2 pg/mL for zearalenone, 0.26 ng/mL for fumonisin B1, 0.11 ng/mL for deoxynivalenol, 15.7 pg/mL for ochratoxin A, and 8.6 pg/mL for T-2 toxin, respectively. The spiking experiment showed high accuracy with recovery of 78.9-106.2 % and satisfactory assay precision with the coefficient of variations below 16 %. Moreover, this assay can be completed in less than 20 min, and its detection results were consistent with that of liquid chromatography-mass spectrometry. Therefore, the developed SERS-based lateral flow immunosensor is a promising approach for mycotoxin detection in the field.