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      Intensified biochip system using chemiluminescence for the detection of Bacillus globigii spores

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          Abstract

          This paper reports the first intensified biochip system for chemiluminescence detection and the feasibility of using this system for the analysis of biological warfare agents is demonstrated. An enzyme-linked immunosorbent assay targeting Bacillus globigii spores, a surrogate species for Bacillus anthracis, using a chemiluminescent alkaline phosphatase substrate is combined with a compact intensified biochip detection system. The enzymatic amplification was found to be an attractive method for detection of low spore concentrations when combined with the intensified biochip device. This system was capable of detecting approximately 1 × 10 5 Bacillus globigii spores. Moreover, the chemiluminescence method, combined with the self-contained biochip design, allows for a simple, compact system that does not require laser excitation and is readily adaptable to field use.

          Figure

          Schematic diagram of the miniature biochip detection system

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          Most cited references23

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          Anthrax as a Biological Weapon, 2002

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            Applications of the luminol chemiluminescent reaction in analytical chemistry.

            This critical review discusses the results published between 2000 and 2005 on the development of analytical systems based on the luminol chemiluminescent and electrochemiluminescent reactions. An increasing number of non-specific detection systems based on the enhancing, inhibiting or catalysing effect of a large range of compounds have been published. Possible detected compounds and their concomitant presence in samples are discussed. Chemiluminescent and electrochemiluminescent reactions were also found to merge in biochip and microarray development as a possible substitute to the well-established but hardly quantitative fluorescent detections.
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              Chemiluminescence detection for hybridization assays on the flow-thru chip, a three-dimensional microchannel biochip.

              Chemiluminescence (CL) detection is seldom used in two-dimensional solid support microarray platforms because adequate sensitivity and spatial resolution is difficult to achieve. The three-dimensional ordered microchannels of the Flow-thru Chip increase both the sensitivity and spatial resolution required for quantitative CL measurements on microarrays. Enzyme-catalyzed CL reactions for the detection of hybridizations on microchannel glass were imaged using a CCD camera. Signal uniformity, sensitivity, and dynamic range of the detection method were determined. The relative standard deviation of signal intensities across an array of 64 spots was 8.1%. A detection limit of 250 amol of target with a linear dynamic range of 3 orders of magnitude was obtained for a 3-h assay. Similar to two-color fluorescence measurements, multiple enzyme labels were employed to demonstrate two-channel chemiluminescence. A unique method for measuring the relaxation time of a chemiluminescent species is also described.
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                Author and article information

                Contributors
                dimitra.stratiscullum1@us.army.mil
                tuan.vodinh@duke.edu
                Journal
                Anal Bioanal Chem
                Analytical and Bioanalytical Chemistry
                Springer-Verlag (Berlin/Heidelberg )
                1618-2642
                1618-2650
                27 January 2008
                July 2008
                : 391
                : 5
                : 1655-1660
                Affiliations
                [1 ]US Army Research Laboratory, AMSRD-ARL-SE-EO, 2800 Powder Mill Road, Adelphi, MD 20783 USA
                [2 ]Fitzpatrick Institute for Photonics, Departments of Biomedical Engineering and Chemistry, Duke University, 2589 Fitzpatrick Center, Durham, NC 27708 USA
                [3 ]Department of Physics and Astronomy, The University of Mississippi, 1025 NCPA, Oxford, MS 38677 USA
                Article
                1835
                10.1007/s00216-008-1835-0
                2467534
                18224472
                105ebd35-b205-42b5-853f-a4222ee91df0
                © Springer-Verlag 2008
                History
                : 30 November 2007
                : 21 December 2007
                : 7 January 2008
                Categories
                Short Communication
                Custom metadata
                © Springer-Verlag 2008

                Analytical chemistry
                bioluminescence,bacillus globigii,chemiluminescence,intensifier,spectroscopy,biosensors immunoassays,elisa,spores,biochips

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