Jump to main content
Jump to site search

Issue 12, 2008
Previous Article Next Article

Integrated microfluidic tmRNA purification and real-time NASBA device for molecular diagnostics

Author affiliations

Abstract

We demonstrate the first integrated microfluidic tmRNA purification and nucleic acid sequence-based amplification (NASBA) device incorporating real-time detection. The real-time amplification and detection step produces pathogen-specific response in < 3 min from the chip-purified RNA from 100 lysed bacteria. On-chip RNA purification uses a new silica bead immobilization method. On-chip amplification uses custom-designed high-selectivity primers and real-time detection uses molecular beacon fluorescent probe technology; both are integrated on-chip with NASBA. Present in all bacteria, tmRNA (10Sa RNA) includes organism-specific identification sequences, exhibits unusually high stability relative to mRNA, and has high copy number per organism; the latter two factors improve the limit of detection, accelerate time-to-positive response, and suit this approach ideally to the detection of small numbers of bacteria. Device efficacy was demonstrated by integrated on-chip purification, amplification, and real-time detection of 100 E. coli bacteria in 100 µL of crude lysate in under 30 min for the entire process.

Graphical abstract: Integrated microfluidic tmRNA purification and real-time NASBA device for molecular diagnostics

Back to tab navigation

Supplementary files

Publication details

The article was received on 21 Jul 2008, accepted on 17 Sep 2008 and first published on 24 Oct 2008


Article type: Paper
DOI: 10.1039/B812515E
Citation: Lab Chip, 2008,8, 2071-2078
  •   Request permissions

    Integrated microfluidic tmRNA purification and real-time NASBA device for molecular diagnostics

    I. K. Dimov, J. L. Garcia-Cordero, J. O'Grady, C. R. Poulsen, C. Viguier, L. Kent, P. Daly, B. Lincoln, M. Maher, R. O'Kennedy, T. J. Smith, A. J. Ricco and L. P. Lee, Lab Chip, 2008, 8, 2071
    DOI: 10.1039/B812515E

Search articles by author

Spotlight

Advertisements