Enzyme-linked immunosorbent-assay for Deoxynivalenol (DON)

Area of Science:

• Food Safety
• Analytical Chemistry
• Mycotoxicology

Background:

• Deoxynivalenol (DON) is a prevalent trichothecene mycotoxin contaminating wheat and barley globally.
• Fusarium Head Blight, caused by Fusarium graminearum, leads to significant DON contamination in cereal crops.
• Accurate detection of DON is crucial due to health risks associated with its ingestion.

Purpose of the Study:

• To develop a rapid, sensitive, and reliable method for the identification and quantification of DON in wheat.
• To establish an indirect competitive enzyme-linked immunosorbent assay (ELISA) for DON detection.

Main Methods:

• Conjugation of 3-O-Hemisuccinyl-DON (3HS-DON) to Bovine serum albumin (BSA) and Ovalbumin (OVA) to generate DON-specific antibodies.
• Optimization of antibody and antigen concentrations for indirect competitive ELISA (1/1600 and 1/1500, respectively).
• Evaluation of assay performance using different extraction buffers and assessment of cross-reactivity with related trichothecenes.

Main Results:

• The developed indirect competitive ELISA demonstrated optimal performance with specific antibody and antigen concentrations.
• The assay showed good recovery rates (82-93%) for DON in grain samples.
• A linear detection range for DON in grains was established between 0.01 and 100 μg/mL, with minimal methanol required in the extraction buffer.

Conclusions:

• The indirect competitive ELISA is a suitable method for the accurate and sensitive detection of DON in wheat.
• The assay is practical for routine analysis of DON contamination in grain samples.
• This method contributes to improved food safety by enabling effective monitoring of mycotoxin levels.