Type

Journal Article

Authors

Luke P. Lee
Antonio J. Ricco
Terry J Smith
Richard O'Kennedy
Majella Maher
Bryan Lincoln
Paul Daly
Lorcan Kent
Caroline Viguier
Claus R Poulsen
and 3 others

Subjects

Microbiology

Topics
microfluidics diagnostic techniques and procedures instrumentation escherichia coli tmrna self sustained sequence replication chemistry rna bacterial

Integrated microfluidic tmRNA purification and real-time NASBA device for molecular diagnostics. (2008)

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 microL of crude lysate in under 30 min for the entire process.
Collections Ireland -> Dublin City University -> PubMed

Full list of authors on original publication

Luke P. Lee, Antonio J. Ricco, Terry J Smith, Richard O'Kennedy, Majella Maher, Bryan Lincoln, Paul Daly, Lorcan Kent, Caroline Viguier, Claus R Poulsen and 3 others

Experts in our system

1
Antonio J. Ricco
Dublin City University
Total Publications: 23
 
2
Terry Smith
National University of Ireland Galway
Total Publications: 43
 
3
R O'Kennedy
Dublin City University
Total Publications: 197