Jump to main content
Jump to site search

Issue 21, 2011
Previous Article Next Article

Miniaturized bead-beating device to automate full DNA sample preparation processes for Gram-positive bacteria

Author affiliations

Abstract

We have developed a miniaturized bead-beating device to automate nucleic acids extraction from Gram-positive bacteria for molecular diagnostics. The microfluidic device was fabricated by sandwiching a monolithic flexible polydimethylsiloxane (PDMS) membrane between two glass wafers (i.e., glass–PDMS–glass), which acted as an actuator for bead collision via its pneumatic vibration without additional lysis equipment. The Gram-positive bacteria, S. aureus and methicillin-resistant S. aureus, were captured on surface-modified glass beads from 1 mL of initial sample solution and in situ lyzed by bead-beating operation. Then, 10 μL or 20 μL of bacterial DNA solution was eluted and amplified successfully by real-time PCR. It was found that liquid volume fraction played a crucial role in determining the cell lysis efficiency in a confined chamber by facilitating membrane deflection and bead motion. The miniaturized bead-beating operation disrupted most of S. aureus within 3 min, which turned out to be as efficient as the conventional benchtop vortexing machine or the enzyme-based lysis technique. The effective cell concentration was significantly enhanced with the reduction of initial sample volume by 50 or 100 times. Combination of such analyte enrichment and in situ bead-beating lysis provided an excellent PCR detection sensitivity amounting to ca. 46 CFU even for the Gram-positive bacteria. The proposed bead-beating microdevice is potentially useful as a nucleic acid extraction method toward a PCR-based sample-to-answer system.

Graphical abstract: Miniaturized bead-beating device to automate full DNA sample preparation processes for Gram-positive bacteria

Back to tab navigation

Supplementary files

Publication details

The article was received on 27 Jul 2011, accepted on 15 Aug 2011 and first published on 14 Sep 2011


Article type: Paper
DOI: 10.1039/C1LC20692C
Citation: Lab Chip, 2011,11, 3649-3655
  •   Request permissions

    Miniaturized bead-beating device to automate full DNA sample preparation processes for Gram-positive bacteria

    K. Hwang, S. H. Kwon, S. Jung, H. Lim, W. Jung, C. Park, J. Kim, K. Suh and N. Huh, Lab Chip, 2011, 11, 3649
    DOI: 10.1039/C1LC20692C

Search articles by author

Spotlight

Advertisements