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Fiber-Optic Biosensor Employing Alexa-Fluor Conjugated Antibody for Detection of Escherichia coli O157:H7 from Ground Beef in Four Hours

Fiber optic biosensor has a great potential to meet the need for rapid, sensitive, and real-time microbial detection systems. We developed an antibody-based fiber-optic biosensor to rapidly detect low levels of Escherichia coli O157:H7 cells in ground beef. The principle of the sensor is a sandwich...

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Detalles Bibliográficos
Autores principales: Geng, Tao, Uknalis, Joe, Tu, Su-I, Bhunia, Arun K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International (MDPI) 2006
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3926523/
Descripción
Sumario:Fiber optic biosensor has a great potential to meet the need for rapid, sensitive, and real-time microbial detection systems. We developed an antibody-based fiber-optic biosensor to rapidly detect low levels of Escherichia coli O157:H7 cells in ground beef. The principle of the sensor is a sandwich immunoassay using an antibody which is specific for E. coli O157:H7. A polyclonal antibody was first immobilized on polystyrene fiber waveguides through a biotin-streptavidin reaction that served as a capture antibody. An Alexa Fluor 647 dye-labeled antibody to E. coli O157:H7 was used to detect cells and generate a specific fluorescent signal, which was acquired by launching a 635 nm laser-light from an Analyte-2000. Fluorescent molecules within several hundred nanometers of the fiber were excited by an evanescent wave, and a portion of the emission light from fluorescent dye transmitted by the fiber and collected by a photodetector at wavelengths of 670 to 710 nm quantitatively. This immunosensor was specific for E. coli O157:H7 compared with multiple other foodborne bacteria. In addition, the biosensor was able to detect as low as 10(3) CFU/ml pure cultured E. coli O157:H7 cells grown in culture broth. Artificially inoculated E. coli O157:H7 at concentration of 1 CFU/ml in ground beef could be detected by this method after only 4 hours of enrichment.