An electrochemiluminescence assay for the detection of bio threat agents in selected food matrices and in the screening of Clostridium botulinum outbreak strains associated with type A botulism

J Sci Food Agric. 2014 Mar 15;94(4):707-12. doi: 10.1002/jsfa.6310. Epub 2013 Aug 12.

Abstract

Background: Specific screening methods for complex food matrices are needed that enable unambiguous and sensitive detection of bio threat agents (BTAs) such as Bacillus anthracis spores and microbial toxins (e.g. staphylococcal enterotoxin B (SEB) and clostridial botulinum neurotoxins (BoNTs)). The present study describes an image-based 96-well Meso Scale Discovery (MSD) electrochemiluminescence (ECL) assay for simultaneous detection of BTAs in dairy milk products.

Results: The limit of detection of this ECL assay is 40 pg mL⁻¹ for BoNT/A complex, 10 pg mL⁻¹ for SEB and 40000 CFU mL⁻¹ for Bacillus anthracis spores in dairy milk products. The ECL assay was successfully applied to screen type A Clostridium botulinum outbreak strains.

Conclusion: The results of the study indicate that this ECL assay is very sensitive, rapid (<6 h) and multiplex in nature. The ECL assay has potential for use as an in vitro screening method for BTAs over other comparable immunoassays.

Keywords: Bacillus anthracis; Clostridium botulinum; bio threat agents; botulinum neurotoxins; botulism; electrochemiluminescene; spores; staphylococcal enterotoxin B.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.
  • Validation Study

MeSH terms

  • Antigens, Bacterial / analysis
  • Antigens, Bacterial / chemistry
  • Antigens, Bacterial / metabolism
  • Bacillus anthracis / growth & development
  • Bacillus anthracis / isolation & purification
  • Bacillus anthracis / physiology
  • Bacterial Toxins / analysis*
  • Bacterial Toxins / chemistry
  • Bacterial Toxins / metabolism
  • Botulinum Toxins, Type A / analysis
  • Botulinum Toxins, Type A / chemistry
  • Botulinum Toxins, Type A / metabolism
  • Botulism / epidemiology
  • Botulism / microbiology
  • Botulism / prevention & control
  • Clostridium botulinum type A / growth & development
  • Clostridium botulinum type A / isolation & purification*
  • Clostridium botulinum type A / metabolism
  • Colony Count, Microbial
  • Dairy Products / adverse effects
  • Dairy Products / analysis*
  • Dairy Products / microbiology
  • Disease Outbreaks / prevention & control
  • Electrochemical Techniques
  • Enterotoxins / analysis
  • Enterotoxins / chemistry
  • Enterotoxins / metabolism
  • Food Contamination*
  • Food Inspection / methods*
  • Food Microbiology
  • Foodborne Diseases / etiology
  • Foodborne Diseases / microbiology
  • Foodborne Diseases / prevention & control*
  • Humans
  • Limit of Detection
  • Luminescence*
  • Luminescent Measurements
  • Spores, Bacterial / isolation & purification
  • United States
  • United States Food and Drug Administration

Substances

  • Antigens, Bacterial
  • Bacterial Toxins
  • Enterotoxins
  • anthrax toxin
  • enterotoxin B, staphylococcal
  • Botulinum Toxins, Type A