Fiber-Optic Biosensor Employing Alexa-Fluor Conjugated Antibody for Detection of Escherichia coli O157H7 from Ground Beef in Four Hours.docVIP

Sensors 2006, 6, 796-807 sensors ISSN 1424-8220 ? 2006 by MDPI /sensors Full Research Paper Fiber-Optic Biosensor Employing Alexa-Fluor Conjugated Antibody for Detection of Escherichia coli O157:H7 from Ground Beef in Four Hours Tao Geng 1, Joe Uknalis 2, Su-I Tu 2 and Arun K. Bhunia 1,* 1Molecular Food Microbiology Laboratory, Department of Food Science, Purdue University, 745 Agriculture Mall Dr., West Lafayette, Indiana 47907, USA 2United States Department of Agriculture, Agricultural Research Service, NAA Eastern Regional Research Center, 600 E. Mermaid Lane, Wyndmoor, PA 19038, USA * Author to whom correspondence should be addressed; E-mail: Bhunia@; Tel. (765) 494- 5443 Received: 3 February 2006 / Accepted 12 April 2006 / Published: 19 August 2006 Abstract: 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 wa