BROWSE

Related Researcher

Author

Soper, Steven A.
Soper Research Group
Research Interests
  • Micro- and nano-fabrication

ITEM VIEW & DOWNLOAD

Ligase Detection Reaction Generation of Reverse Molecular Beacons for Near Real-Time Analysis of Bacterial Pathogens Using Single-Pair Fluorescence Resonance Energy Transfer and a Cyclic Olefin Copolymer Microfluidic Chip

Cited 16 times inthomson ciCited 16 times inthomson ci
Title
Ligase Detection Reaction Generation of Reverse Molecular Beacons for Near Real-Time Analysis of Bacterial Pathogens Using Single-Pair Fluorescence Resonance Energy Transfer and a Cyclic Olefin Copolymer Microfluidic Chip
Author
Peng, ZhiyongSoper, Steven A.Pingle, Maneesh R.Barany, FrancisDavis, Lloyd M.
Keywords
16S rRNA gene; Bacterial pathogens; Close proximity; Continuous flows; Cyclic Olefin Copolymers; Fluorescence resonance energy transfer; Innovative strategies; Laser induced fluorescence; Ligase detection reactions; Micro-fluidic devices; Microfluidic chip; Molecular beacon; On chips; Pathogenic bacterium; Potential threats; Real time analysis; Single-molecule detection; Single-molecule photons; Specific primers; Target sequences; Thermal cycle
Issue Date
201012
Publisher
AMER CHEMICAL SOC
Citation
ANALYTICAL CHEMISTRY, v.82, no.23, pp.9727 - 9735
Abstract
Detection of pathogenic bacteria and viruses require strategies that can signal the presence of these targets in near real-time due to the potential threats created by rapid dissemination into water and/or food supplies In this paper, we report an innovative strategy that can rapidly detect bacterial pathogens using reporter sequences found in their genome without requiring polymerase chain reaction (PCR) A pair of strain-specific primers was designed based on the 168 rRNA gene and were end-labeled with a donor (Cy5) or acceptor (Cy5 5) dye In the presence of the target bacterium, the primers were joined using a ligase detection reaction (LDR) only when the primers were completely complementary to the target sequence to form a reverse molecular beacon (rMB), thus bringing Cy5 (donor) and Cy5 5 (acceptor) into close proximity to allow fluorescence resonance energy transfer (FRET) to occur These rMBs were subsequently analyzed using single molecule detection of the FRET pairs (single-pair FRET, spFRET) The LDR was performed using a continuous flow thermal cycling process configured in a cyclic olefin copolymer (COC) microfluidic device using either 2 or 20 thermal cycles Single molecule photon bursts from the resulting rMBs were detected on chip and registered using a simple laser-induced fluorescence (LIF) instrument. The spFRET signatures from the target pathogens were reported in as little as 2 6 mm using spFRET.
URI
Go to Link
DOI
http://dx.doi.org/10.1021/ac101843n
ISSN
0003-2700
Appears in Collections:
ECHE_Journal Papers

find_unist can give you direct access to the published full text of this article. (UNISTARs only)

Show full item record

qr_code

  • mendeley

    citeulike

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

MENU