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

Miyajima K, Koshida T, Arakawa T, Kudo H, Saito H, Yano K, Mitsubayashi K - Biosensors (Basel) (2013)

Bottom Line: The measurement for each sample was completed within 12 min.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.

View Article: PubMed Central - PubMed

Affiliation: 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; E-Mail: miya.bdi@tmd.ac.jp ; 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; E-Mails: arakawa.bdi@tmd.ac.jp (T.A.); kudo.bdi@tmd.ac.jp (H.K.).

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 10(3) to 10(7) 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.

No MeSH data available.


Fiber-optic fluoroimmunoassay system consisting of a flow-through cell. The flow cell is made from poly(methyl methacrylate) and the capacity is 350 μL.
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biosensors-03-00120-f002: Fiber-optic fluoroimmunoassay system consisting of a flow-through cell. The flow cell is made from poly(methyl methacrylate) and the capacity is 350 μL.

Mentions: The fluorometric assay system (PU6102) and polystyrene optical fiber probes (4 cm in length and 0.78 mm in diameter) were contributed by Canon Chemicals Inc. One end of the probe had a lens to effectively collect the excitation light and to guide it to the probe. Figure 1 shows the principle of the fluoroimmunoassay system for E. coli O157:H7. The measurement was performed using the customized PU6102 fluorometric assay system with the flow cell. The assay principle is based on a sandwich immunoassay, using optical fiber probes coated with capture antibodies and Cy5-labeled antibodies for fluorometric detection. The excitation light (λ = 650 nm) from a laser diode (LD) is coupled to the optical fiber probe, and the light is transmitted in the probe by total internal reflection. Then the Cy5 florescent molecules near the probe surface are excited by an evanescent light at the surface of the probe. E. coli O157:H7 is measured as fluorescence of Cy5 using a photodiode. Figure 2 shows the appearance of the customized PU6102 system and specially designed flow cell. The flow cell is made from poly(methyl methacrylate) and the capacity is 350 μL (except paths of flow).


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

Miyajima K, Koshida T, Arakawa T, Kudo H, Saito H, Yano K, Mitsubayashi K - Biosensors (Basel) (2013)

Fiber-optic fluoroimmunoassay system consisting of a flow-through cell. The flow cell is made from poly(methyl methacrylate) and the capacity is 350 μL.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4263595&req=5

biosensors-03-00120-f002: Fiber-optic fluoroimmunoassay system consisting of a flow-through cell. The flow cell is made from poly(methyl methacrylate) and the capacity is 350 μL.
Mentions: The fluorometric assay system (PU6102) and polystyrene optical fiber probes (4 cm in length and 0.78 mm in diameter) were contributed by Canon Chemicals Inc. One end of the probe had a lens to effectively collect the excitation light and to guide it to the probe. Figure 1 shows the principle of the fluoroimmunoassay system for E. coli O157:H7. The measurement was performed using the customized PU6102 fluorometric assay system with the flow cell. The assay principle is based on a sandwich immunoassay, using optical fiber probes coated with capture antibodies and Cy5-labeled antibodies for fluorometric detection. The excitation light (λ = 650 nm) from a laser diode (LD) is coupled to the optical fiber probe, and the light is transmitted in the probe by total internal reflection. Then the Cy5 florescent molecules near the probe surface are excited by an evanescent light at the surface of the probe. E. coli O157:H7 is measured as fluorescence of Cy5 using a photodiode. Figure 2 shows the appearance of the customized PU6102 system and specially designed flow cell. The flow cell is made from poly(methyl methacrylate) and the capacity is 350 μL (except paths of flow).

Bottom Line: The measurement for each sample was completed within 12 min.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.

View Article: PubMed Central - PubMed

Affiliation: 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; E-Mail: miya.bdi@tmd.ac.jp ; 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; E-Mails: arakawa.bdi@tmd.ac.jp (T.A.); kudo.bdi@tmd.ac.jp (H.K.).

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 10(3) to 10(7) 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.

No MeSH data available.