APPLICATIONS OF TIME‐RESOLVED FLUOROIMMUNOASSAY TO DETECT MAGNETIC BEAD CAPTURED <i>ESCHERICHIA COLI</i> O157:H7

Tu, Shu‐I; Golden, M.H.; Andreotti, Peter E.; Yu, Linda; Irwin, Peter L.

doi:10.1111/j.1745-4581.2001.tb00231.x

Cited by 23 publications

(25 citation statements)

References 21 publications

(10 reference statements)

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“…Thus, our current process is much simpler in operation. The current approach appears to be even faster than the time-resolved fluoroimmunoassay (TRF) that requires a 4.5 h enrichment for detection of 1 CFU/g of E. coli O157:H7 in ground beef [26, 27]. …”

Section: Resultsmentioning

confidence: 99%

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

Geng

Uknalis

et al. 2006

Sensors

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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 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.

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Section: Resultsmentioning

confidence: 99%

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

Geng

Uknalis

et al. 2006

Sensors

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“…In fluorescence-based immunoassays, the common separation step can be simplified by the aid of magnetic particles [21,27,48,199,200]. Kits for LuMaPs-based immunoassays including necessary separators are commercially available.…”

Section: Fluorescence-based Immunoassaysmentioning

confidence: 99%

Luminescent magnetic particles: structures, syntheses, multimodal imaging, and analytical applications

Mistlberger

Klimant

2010

Bioanal Rev

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Luminescent magnetic particles (LuMaPs) are attractive tools for life science applications such as multimodal imaging, analyte monitoring, nanotherapeutics, and combinations thereof. LuMaPs consist of at least one magnetic and one luminescent component which often are incorporated in a (polymeric) matrix. A large variety of materials do exist for the components that make up LuMaPs. However, a smart selection and combination is required for achieving useful tools. While the magnetic component mainly influences the response to a magnetic field, the luminophore can act as label, sensor, or therapeutic agent. The matrix fulfills tasks such as stabilizing the luminophore and magnet, carrying useful functional groups on its surface, or hosting smart drug delivery systems. Surface modifications with targeting ligands can further improve the applicability of LuMaPs, for example in biomedicine. This review provides an overview on LuMaPs with respect to the materials used and to its structures. Routes toward LuMaPs are outlined, and potential applications are discussed.

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“…In an effort to provide both rapid and sensitive detection of pathogens once they enter the food chain, we have combined the immunomagnetic capture and separation of target organisms with a sensitive method of fluorescence detection-time-resolved fluorescence (TRF; Tu et al 2001). TRF utilizes lanthanide chelate labels that are bound to a secondary antibody specific to the target of interest.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Detection of Escherichia coli O157:H7 and Salmonella Typhimurium: The Use of Magnetic Beads Conjugated with Multiple Capture Antibodies

Reed

Gehring

et al. 2010

Food Anal. Methods

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Streptavidin-coated magnetic beads were conjugated with biotinylated capture antibodies to both Escherichia coli O157:H7 and Salmonella Typhimurium to form multipathogen capture immunomagnetic beads (IMB-M). The efficacy of these beads was investigated in both pure and mixed culture suspensions, as well as in inoculated spinach and ground beef. Using dual-label time-resolved fluorescence, a sandwich immunoassay consisting of europium-and samarium-labeled detection antibodies was used to measure the capture of E. coli and Salmonella, respectively. IMB-M was just as effective as the mixture of IMB against E. coli and Salmonella in capturing both organisms in pure culture and E. coli in mixed culture. Capture of Salmonella in mixed culture by IMB-M resulted in lower fluorescent signals, but comparable detection limit. In inoculated food samples, IMB-M was just as effective as the mixture of IMB specific for both target organisms in detecting 1 cfu/g of E. coli and 100 cfu/g of Salmonella after a 6-h enrichment at 37°C. Multi-pathogen capture IMB have the potential to be convenient and important tools for multi-pathogen detection.

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APPLICATIONS OF TIME‐RESOLVED FLUOROIMMUNOASSAY TO DETECT MAGNETIC BEAD CAPTURED ESCHERICHIA COLI O157:H7

Cited by 23 publications

References 21 publications

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

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

Luminescent magnetic particles: structures, syntheses, multimodal imaging, and analytical applications

Simultaneous Detection of Escherichia coli O157:H7 and Salmonella Typhimurium: The Use of Magnetic Beads Conjugated with Multiple Capture Antibodies

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