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Soper, Steven A.
Soper Research Group
Research Interests
  • Micro- and nano-fabrication
  • Lab-on-a-chip
  • Polymeric Microfluidic Devices

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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

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dc.contributor.author Peng, Zhiyong ko
dc.contributor.author Soper, Steven A. ko
dc.contributor.author Pingle, Maneesh R. ko
dc.contributor.author Barany, Francis ko
dc.contributor.author Davis, Lloyd M. ko
dc.date.available 2014-05-07T04:38:48Z -
dc.date.created 2013-06-20 ko
dc.date.issued 2010-12 -
dc.identifier.citation ANALYTICAL CHEMISTRY, v.82, no.23, pp.9727 - 9735 ko
dc.identifier.issn 0003-2700 ko
dc.identifier.uri https://scholarworks.unist.ac.kr/handle/201301/4489 -
dc.identifier.uri http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=78649701493 ko
dc.description.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. ko
dc.description.statementofresponsibility close -
dc.language ENG ko
dc.publisher AMER CHEMICAL SOC ko
dc.subject 16S rRNA gene ko
dc.subject Bacterial pathogens ko
dc.subject Close proximity ko
dc.subject Continuous flows ko
dc.subject Cyclic Olefin Copolymers ko
dc.subject Fluorescence resonance energy transfer ko
dc.subject Innovative strategies ko
dc.subject Laser induced fluorescence ko
dc.subject Ligase detection reactions ko
dc.subject Micro-fluidic devices ko
dc.subject Microfluidic chip ko
dc.subject Molecular beacon ko
dc.subject On chips ko
dc.subject Pathogenic bacterium ko
dc.subject Potential threats ko
dc.subject Real time analysis ko
dc.subject Single-molecule detection ko
dc.subject Single-molecule photons ko
dc.subject Specific primers ko
dc.subject Target sequences ko
dc.subject Thermal cycle ko
dc.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 ko
dc.type ARTICLE ko
dc.identifier.scopusid 2-s2.0-78649701493 ko
dc.identifier.wosid 000284668600021 ko
dc.type.rims ART ko
dc.description.wostc 16 *
dc.description.scopustc 16 *
dc.date.tcdate 2014-10-18 *
dc.date.scptcdate 2014-07-12 *
dc.identifier.doi 10.1021/ac101843n ko
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