Signal amplification using colloidal gold in a biolayer interferometry-based immunosensor for the mycotoxin deoxynivalenol

Abstract

Deoxynivalenol (DON) is a toxin produced by certain species of Fusarium fungi that can infest wheat, barley and corn. The fungi cause diseases in crops worldwide and some of the secondary metabolites, such as DON, can adversely affect animal health and food safety. To monitor DON in wheat rapidly, a biosensor using the principle of biolayer interferometry (BLI) was developed. The signal from the sensor was substantially amplified through the use of a primary antibody–colloidal gold conjugate. The amplification was much greater in the presence of wheat matrix than in buffered solution, suggesting matrix components may have contributed to the enhancement. The improved signal provided by the amplification allowed for the development of rapid qualitative and quantitative assays. The limit of detection of the method was 0.09 mg kg⁻¹; the limit of quantitation was 0.35 mg kg⁻¹. Recovery from wheat spiked over the range from 0.2 to 5 mg kg⁻¹ averaged 103% (RSD = 12%). The quantitative assay compared favourably (r ²= 0.9698) with a reference chromatographic method for 40 naturally contaminated wheats. The qualitative assay was able to classify accurately the same group of 40 samples as either above or below a 0.5 mg kg⁻¹ threshold. These results suggest that the BLI technique can be used to measure DON in wheat rapidly.

Keywords:

• screening – immunoassays
• screening – biosensor
• mycotoxins – trichothecenes
• trichothecenes
• cereals and grain

Acknowledgements

The author wishes to thank Mr John Bobell, USDA-ARS-NCAUR, Peoria, IL, USA, for his exceptional technical assistance. Disclaimer: The mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture (USDA). The USDA is an equal opportunity provider and employer.