Source:http://linkedlifedata.com/resource/pubmed/id/15167797
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
11
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pubmed:dateCreated |
2004-5-28
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pubmed:abstractText |
An integrated portable genetic analysis microsystem including PCR amplification and capillary electrophoretic (CE) analysis coupled with a compact instrument for electrical control and laser-excited fluorescence detection has been developed. The microdevice contains microfabricated heaters, temperature sensors, and membrane valves to provide controlled sample positioning and immobilization in 200-nL PCR chambers. The instrument incorporates a solid-state laser and confocal fluorescence detection optics, electronics for sensing and powering the PCR reactor, and high-voltage power supplies for conducting CE separations. The fluorescein-labeled PCR products are amplified and electrophoretically analyzed in a gel-filled microchannel in <10 min. We demonstrate the utility of this instrument by performing pathogen detection and genotyping directly from whole Escherichia coli and Staphylococcus aureus cells. The E. coli detection assay consists of a triplex PCR amplification targeting genes that encode 16S ribosomal RNA, the fliC flagellar antigen, and the sltI shigatoxin. Serial dilution demonstrates a limit of detection of 2-3 bacterial cells. The S. aureus assay uses a femA marker to identify cells as S. aureus and a mecA marker to probe for methicillin resistance. This integrated portable genomic analysis microsystem demonstrates the feasibility of performing rapid high-quality detection of pathogens and their antimicrobial drug resistance.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:status |
MEDLINE
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pubmed:month |
Jun
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pubmed:issn |
0003-2700
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
1
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pubmed:volume |
76
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
3162-70
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:15167797-Animals,
pubmed-meshheading:15167797-Bacteria,
pubmed-meshheading:15167797-Bacterial Infections,
pubmed-meshheading:15167797-Base Sequence,
pubmed-meshheading:15167797-Electrophoresis, Capillary,
pubmed-meshheading:15167797-Escherichia coli Infections,
pubmed-meshheading:15167797-Molecular Sequence Data,
pubmed-meshheading:15167797-Oligonucleotide Array Sequence Analysis,
pubmed-meshheading:15167797-Polymerase Chain Reaction,
pubmed-meshheading:15167797-Staphylococcal Infections
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pubmed:year |
2004
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pubmed:articleTitle |
Integrated portable genetic analysis microsystem for pathogen/infectious disease detection.
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pubmed:affiliation |
Chemistry Department, University of California, Berkeley, California 94720, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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