A flow cytometry based competitive fluorescent microsphere immunoassay (CFIA) system for detecting up to six mycotoxins

J Immunol Methods. 2012 Oct 31;384(1-2):71-80. doi: 10.1016/j.jim.2012.07.010. Epub 2012 Jul 25.

Abstract

Background: Exposure to multiple mycotoxins through the food chain represents a major potential health hazard to both humans and livestock. They can cause a variety of severe acute as well as chronic diseases. Eliminating mycotoxins from various grain crops is a global health priority. According to the Food and Agriculture Organization (FAO), world food production needs to double by 2050. Innovative solutions will be required to sustain toxin free grain supplies worldwide.

Methods: A competitive flow cytometry based multiplexed assay with fluorescent microspheres has been developed. The new multiplexed method can analyze simultaneously any one or all six major mycotoxins. They include: Ochratoxin A (OTA), Aflatoxin B1 (AFB1), Fumonisin B1 (FB1), T-2 toxin (T-2), Deoxynivalenol (DON) and Zearalenone (ZEA), which are all potential human health hazards. The CFIA described here includes a simplified single extraction step for mycotoxins from specimens and a comprehensive post acquisition software module. The new assay system was developed with a FACSArray™ BD Bioanalyzer flow cytometer (BD Biosciences, Belgium).

Results: The CFIA performs favourably when compared to commercial ELISA. Sensitivity range with CFIA increased between 13% and 100% with an average improvement of 50% for the six mycotoxins.

Conclusions: The multiplexed assay presented here has the unique capacity to quantify up to six mycotoxins simultaneously from a single specimen extraction. CFIA's poly-mycotoxin detection sensitivity exceeds standard ELISA. CFIA may be part of a comprehensive assay system that will provide reliable and effective safeguard for agricultural commodities to be free of mycotoxins.

MeSH terms

  • Aflatoxin B1 / analysis
  • Aflatoxin B1 / chemistry
  • Aflatoxin B1 / immunology
  • Animals
  • Antibodies, Fungal / immunology
  • Enzyme-Linked Immunosorbent Assay / methods
  • Flow Cytometry / methods*
  • Fluorescent Dyes / chemistry
  • Fumonisins / analysis
  • Fumonisins / chemistry
  • Fumonisins / immunology
  • Humans
  • Immunoassay / methods*
  • Mice
  • Microspheres
  • Mycotoxins / analysis*
  • Mycotoxins / chemistry
  • Mycotoxins / immunology*
  • Ochratoxins / analysis
  • Ochratoxins / chemistry
  • Ochratoxins / immunology
  • Reproducibility of Results
  • Sensitivity and Specificity
  • T-2 Toxin / analysis
  • T-2 Toxin / chemistry
  • T-2 Toxin / immunology
  • Trichothecenes / analysis
  • Trichothecenes / chemistry
  • Trichothecenes / immunology
  • Zearalenone / analysis
  • Zearalenone / chemistry
  • Zearalenone / immunology

Substances

  • Antibodies, Fungal
  • Fluorescent Dyes
  • Fumonisins
  • Mycotoxins
  • Ochratoxins
  • Trichothecenes
  • ochratoxin A
  • fumonisin B1
  • Zearalenone
  • Aflatoxin B1
  • T-2 Toxin
  • deoxynivalenol