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Proceedings Paper

Development of a near-real-time procedure to detect Staphylococcus aureus enterotoxin A in military rations
Author(s): Michelle J. Richardson; Arthur Garth Rand; Andre G. Senecal
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Paper Abstract

Using a chemiluminescent fiber optic biosensor and magnetic particles, a simple, sensitive and rapid method to determine Staphylococcus aureus enterotoxin A (SEA) in military ration components was developed. Anti-staphylococcal enterotoxin A (Anti-SEA) was immobilized on magnetic particles and incubated with SEA. The beads were then collected and rinsed on a membrane filter (0.45um). The captured toxin was then selectively labeled with a monoclonal-horseradish peroxidase (POD) conjugate. SEA concentration was detected with a luminometer and a chemiluminescent enhancing reagent. Total assay time was 1.25 hours. Chemiluminescent signal due to nonspecific binding was tested with various blocking agents. Phosphate buffered saline with casein had the lowest background signal. Primary antibody concentration, secondary labeled antibody concentration and chemiluminescent substrate type were also evaluated to optimize signal intensity. The chemiluminescent fiber optic biosensor assay was compared to the Analyte 2000, a commercial fluorescent fiber optic biosensor. This assay consisted of immobilizing Anti-SEA on polystyrene waveguides, and incubating the waveguides with the toxin. The waveguide was incubated with a selectively labeled monoclonal-CY5 Dye conjugate. The sensitivity of chemiluminescent and fluorescent immunoassays were 1 ng, significantly lower than the levels needed to cause illness.

Paper Details

Date Published: 21 February 2002
PDF: 12 pages
Proc. SPIE 4575, Chemical and Biological Early Warning Monitoring for Water, Food, and Ground, (21 February 2002); doi: 10.1117/12.456919
Show Author Affiliations
Michelle J. Richardson, U.S. Army Soldier and Biological Chemical Command and Univ. of Rhode Island (United States)
Arthur Garth Rand, Univ. of Rhode Island (United States)
Andre G. Senecal, U.S. Army Soldier and Biological Chemical Command (United States)


Published in SPIE Proceedings Vol. 4575:
Chemical and Biological Early Warning Monitoring for Water, Food, and Ground
Janet L. Jensen; Larry W. Burggraf, Editor(s)

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