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Electrochemiluminescence DNA sensor array for multiplex detection of biowarfare agents

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Abstract

Development of a fully automated electrochemiluminescence (ECL) DNA assay for multiplex detection of six biowarfare agents is described. Aminated-DNA capture probes were covalently immobilised on activated-carbon electrodes and subsequently hybridised to target strands. Detection was achieved via a sandwich-type assay after Ru(bpy)3 2+-labelled reporter probes were hybridised to the formed probe–target complexes. The assay was performed in an automated microsystem in a custom-designed ECL detection box with integrated fluidics, electronics, and movable photomultiplier detector. The obtained limits of detection were 0.6–1.2 nmol L−1 for six targets ranging from 50 to 122 base pairs in size, with linear range 1–15 nmol L−1. Non-specific adsorption and cross-reactivity were very low. Detection of six targets on a single chip was achieved with subnanomolar detection limits.

A photo of the electrode array containing 3 × 14 carbon working electrodes (WE), a common carbon counter electrode (CE), and Ag/AgCl reference electrode (RE) (top). The detection scheme is indicated for one WE. First, probe sequences were covalently immobilised on the activated-carbon surface, then target strands were introduced, and finally Ru(bpy)3 2+-labelled reporter strands were introduced to complete the sandwich-type hybridisation assay. The 3D graph shows multiplex detection of six different pathogens on a single chip

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Acknowledgments

The authors thank microfluidic ChipShop (http://www.microfluidic-chipshop.com/) for provision of the fluidic chips.

Funding

The research was funded by the European Community’s Seventh Framework Programme MultisenseChip, ID number: FP7/2007-2013.

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Authors and Affiliations

  1. Nanobiotechnology and Bioanalysis Group, INTERFIBIO Consolidated Research Group, Departament d’Enginyeria Quimica, Universitat Rovira i Virgili, Avinguda Països Catalans 26, 43007, Tarragona, Spain

    Anna-Maria Spehar-Délèze, Samuel Dulay & Ciara K. O’Sullivan

  2. Fraunhofer ICT-IMM, Carl-Zeiss-Strasse 18-20, 55129, Mainz, Germany

    Rainer Gransee

  3. iMicroQ, Ferro 6 (nau 7), Poligono Industrial Riu Clar, 43006, Tarragona, Spain

    Sergio Martinez-Montequin & Diego Bejarano-Nosas

  4. Institute of Bacterial Infections and Zoonoses, Friedrich-Loeffler-Institut, Naumburger Str. 96A, 07743, Jena, Germany

    Sandra Julich & Herbert Tomaso

  5. Institució Catalana de Recerca i Estudis Avancats, Passeig Lluís Companys, 23, 08010, Barcelona, Spain

    Ciara K. O’Sullivan

Authors
  1. Anna-Maria Spehar-Délèze
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  2. Rainer Gransee
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  3. Sergio Martinez-Montequin
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  4. Diego Bejarano-Nosas
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  5. Samuel Dulay
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  6. Sandra Julich
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  7. Herbert Tomaso
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  8. Ciara K. O’Sullivan
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Corresponding authors

Correspondence to Anna-Maria Spehar-Délèze or Ciara K. O’Sullivan.

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Spehar-Délèze, AM., Gransee, R., Martinez-Montequin, S. et al. Electrochemiluminescence DNA sensor array for multiplex detection of biowarfare agents. Anal Bioanal Chem 407, 6657–6667 (2015). https://doi.org/10.1007/s00216-015-8831-y

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  • DOI: https://doi.org/10.1007/s00216-015-8831-y

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