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

Michelle M Packard; Maxim Shusteff; Evangelyn C Alocilja

doi:10.3390/bios2040405

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

Biosensors (Basel). 2012 Oct 10;2(4):405-16. doi: 10.3390/bios2040405.

Authors

Michelle M Packard¹, Maxim Shusteff², Evangelyn C Alocilja³

Affiliations

¹ Nanobiosensors Laboratory, Michigan State University, East Lansing, MI 48824, USA.
² Lawrence Livermore National Laboratory, 7000 East Ave., Livermore, CA 94550, USA. shusteff1@llnl.gov.
³ Nanobiosensors Laboratory, Michigan State University, East Lansing, MI 48824, USA. alocilja@msu.edu.

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.