Flow cytometry based rapid duplexed immunoassay for fusarium mycotoxins

Area of Science:

• Food Science and Technology
• Analytical Chemistry
• Biotechnology

Background:

• Enzyme-linked immunosorbent assay (ELISA) is widely used for mycotoxin testing in grain but is inefficient for multiple contaminants.
• Increasing co-occurrence of mycotoxins necessitates more effective detection methods for food safety compliance.
• Current ELISA methods require separate sample preparation and long incubation times for each mycotoxin.

Purpose of the Study:

• To modify a commercial competitive fluorescent microsphere immunoassay (CFIA) kit for point-of-need (PON) applications.
• To develop a multiplexed assay capable of simultaneously detecting multiple mycotoxins in grain.
• To reduce the overall processing time for mycotoxin analysis in food safety testing.

Main Methods:

• Modification of an existing commercial CFIA kit for enhanced utility.
• Development of a single, universal extraction protocol for multiple mycotoxins.
• Investigation of binding kinetics for suspended solid-phase particles in liquid phase.

Main Results:

• The accelerated CFIA protocol achieved over a 60% reduction in total processing time.
• The modified CFIA successfully detected dual mycotoxin contamination simultaneously.
• Binding kinetics were analyzed and supported by theoretical calculations.

Conclusions:

• The developed multiplexed CFIA offers a cost-effective and time-efficient alternative to ELISA for mycotoxin detection.
• This assay is suitable for point-of-need applications in small food processing facilities.
• Future advancements in portable cytometry could enable rapid, multiplexed testing for enhanced food safety.