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Proceedings Paper

Rapid detection of Staphylococcus aureus using a membrane fiber optic biosensor
Author(s): Jianming Ye; Philip E. Pivarnik; Andre G. Senecal; Arthur Garth Rand
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Paper Abstract

A simple, sensitive and rapid chemiluminescent fiber optic biosensor utilizing monoclonal antibodies to S. aureus was developed to detect the pathogen in food. The S. aureus cells were selectively labeled with a monoclonal-horseradish peroxidase (POD) conjugate, collected by membrane filtration, and detected with a luminometer and an enhanced chemiluminescent luminol reagent. Two different diameter membranes, 25 mm and 13 mm, were first tested in a luminometer tube format assay. A hand operated syringe filtration unit was used to capture cells and the membrane was then transferred to a luminometer tube for the chemiluminescent reaction. An improved system utilized a simple but efficient microwell plate vacuum filtration unit with an 8 mm membrane sealed at the bottom of the sample well. The sample was concentrated on the membrane and positioned directly in front of a fiber optic light guide to effectively collect and transmit the signal to the luminometer. Labeling S. aureus in solution proved to be much more effective than on the membrane surface. Using the microwell plate filtration system resulted in less sample handling, better reproducibility, and dramatically reduced assay time. The variability for 25 mm and 13 mm assays were 24.7% and 13.3%, while the microwell plate assay reduced this to 4.0%. The ability of the fiber optic probe to effectively collect the signal meant the sensitivity of the assay was not compromised with smaller membrane and sample size. The sensitivity of the biosensor was 3.8 X 104 CFU/ml, adequate to detect the organism at concentrations lower than the level that could result in food poisoning. The performance of the biosensor was not effected by the food materials and by the presence of other bacteria.

Paper Details

Date Published: 12 January 1999
PDF: 8 pages
Proc. SPIE 3544, Pathogen Detection and Remediation for Safe Eating, (12 January 1999); doi: 10.1117/12.335766
Show Author Affiliations
Jianming Ye, Univ. of Rhode Island (United States)
Philip E. Pivarnik, Univ. of Rhode Island (United States)
Andre G. Senecal, U.S. Army Soldier Systems Command (Natick) (United States)
Arthur Garth Rand, Univ. of Rhode Island (United States)

Published in SPIE Proceedings Vol. 3544:
Pathogen Detection and Remediation for Safe Eating
Yud-Ren Chen, Editor(s)

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