Biosensors 2013, 3(1), 120-131; doi:10.3390/bios3010120
Article

Fiber-Optic Fluoroimmunoassay System with a Flow-Through Cell for Rapid On-Site Determination of Escherichia coli O157:H7 by Monitoring Fluorescence Dynamics

Kumiko Miyajima 1,2 email, Tomoyuki Koshida 3 email, Takahiro Arakawa 2 email, Hiroyuki Kudo 2 email, Hirokazu Saito 4 email, Kazuyoshi Yano 3 email and Kohji Mitsubayashi 1,2,* email
1 Department of Advanced Sciences and Technology for Biomedical Sensors, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549, Japan 2 Department of Biomedical Devices and Instrumentation, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-Surugadai, Chiyoda-ku, Tokyo 101-0062, Japan 3 Graduate School of Bionics, Tokyo University of Technology, 1404-1, Katakura-machi, Hachioji-shi, Tokyo 192-0982, Japan 4 Department of Mechanical Engineering, Tokyo National Collage of Technology, 1220-2 Kunugida-machi, Hachioji-shi, Tokyo 193-0997, Japan
* Author to whom correspondence should be addressed.
Received: 8 January 2013; in revised form: 13 February 2013 / Accepted: 5 March 2013 / Published: 8 March 2013
(This article belongs to the Special Issue Immunosensors 2012)
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Abstract: Dynamic fluoroimmunoassay with a flow-through system using optical fiber probes consisting of polystyrene was developed and applied to a quantitative detection of E. coli O157:H7. The system measures E. coli as fluorescence of sandwich-type immune complexes formed by capture antibodies immobilized on the surface of the probe, E. coli cells, and fluorescently labeled detection antibodies. Excitation was carried out using an evanescent wave from the probe. Resulting fluorescence recoupled into the probe was detected by a photodiode. The assay system was constructed with a flow cell which was available for sequential injection of experimental reagents. In vitro characterization was performed using the flow cell, and the calibration range of E. coli O157:H7 was from 103 to 107 cells/mL. The measurement for each sample was completed within 12 min. Furthermore, it was also possible to estimate the concentrations of E. coli O157:H7 by the increasing rate of fluorescence during binding reaction of detection antibodies to antigens. This minimized the time for measurement down to 6 min. The system is suitable for rapid and direct determination for microorganisms or bacteria in food, clinical, and environmental sources.
Keywords: E. coli O157:H7; fluorescence; immunoassay; optical fiber; flow cell

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Cite This Article

MDPI and ACS Style

Miyajima, K.; Koshida, T.; Arakawa, T.; Kudo, H.; Saito, H.; Yano, K.; Mitsubayashi, K. Fiber-Optic Fluoroimmunoassay System with a Flow-Through Cell for Rapid On-Site Determination of Escherichia coli O157:H7 by Monitoring Fluorescence Dynamics. Biosensors 2013, 3, 120-131.

AMA Style

Miyajima K, Koshida T, Arakawa T, Kudo H, Saito H, Yano K, Mitsubayashi K. Fiber-Optic Fluoroimmunoassay System with a Flow-Through Cell for Rapid On-Site Determination of Escherichia coli O157:H7 by Monitoring Fluorescence Dynamics. Biosensors. 2013; 3(1):120-131.

Chicago/Turabian Style

Miyajima, Kumiko; Koshida, Tomoyuki; Arakawa, Takahiro; Kudo, Hiroyuki; Saito, Hirokazu; Yano, Kazuyoshi; Mitsubayashi, Kohji. 2013. "Fiber-Optic Fluoroimmunoassay System with a Flow-Through Cell for Rapid On-Site Determination of Escherichia coli O157:H7 by Monitoring Fluorescence Dynamics." Biosensors 3, no. 1: 120-131.

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