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Biosensors 2012, 2(4), 405-416; doi:10.3390/bios2040405
Article

Microfluidic-Based Amplification-Free Bacterial DNA Detection by Dielectrophoretic Concentration and Fluorescent Resonance Energy Transfer Assisted in Situ Hybridization (FRET-ISH)

1,2,#, 2
 and 1,*
Received: 20 August 2012; in revised form: 24 September 2012 / Accepted: 8 October 2012 / Published: 10 October 2012
(This article belongs to the Special Issue Nano and Micro DNA/RNA Sensors)
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Abstract: Although real-time PCR (RT-PCR) has become a diagnostic standard for rapid identification of bacterial species, typical methods remain time-intensive due to sample preparation and amplification cycle times. The assay described in this work incorporates on-chip dielectrophoretic capture and concentration of bacterial cells, thermal lysis, cell permeabilization, and nucleic acid denaturation and fluorescence resonance energy transfer assisted in situ hybridization (FRET-ISH) species identification. Combining these techniques leverages the benefits of all of them, allowing identification to be accomplished completely on chip less than thirty minutes after receipt of sample, compared to multiple hours required by traditional RT-PCR and its requisite sample preparation.
Keywords: fluorescence resonance energy transfer assisted in situ hybridization (FRET-ISH); in situ hybridization (ISH); dielectrophoresis (DEP); on-chip diagnostics; lab on a chip; microbial identification fluorescence resonance energy transfer assisted in situ hybridization (FRET-ISH); in situ hybridization (ISH); dielectrophoresis (DEP); on-chip diagnostics; lab on a chip; microbial identification
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Packard, M.M.; Shusteff, M.; Alocilja, E.C. Microfluidic-Based Amplification-Free Bacterial DNA Detection by Dielectrophoretic Concentration and Fluorescent Resonance Energy Transfer Assisted in Situ Hybridization (FRET-ISH). Biosensors 2012, 2, 405-416.

AMA Style

Packard MM, Shusteff M, Alocilja EC. Microfluidic-Based Amplification-Free Bacterial DNA Detection by Dielectrophoretic Concentration and Fluorescent Resonance Energy Transfer Assisted in Situ Hybridization (FRET-ISH). Biosensors. 2012; 2(4):405-416.

Chicago/Turabian Style

Packard, Michelle M.; Shusteff, Maxim; Alocilja, Evangelyn C. 2012. "Microfluidic-Based Amplification-Free Bacterial DNA Detection by Dielectrophoretic Concentration and Fluorescent Resonance Energy Transfer Assisted in Situ Hybridization (FRET-ISH)." Biosensors 2, no. 4: 405-416.

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