| A rapid multiplex assay for nucleic acid-based diagnostics. | |
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MedLine Citation:
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PMID: 20006656 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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We have developed a rapid (under 4 hours), multiplex, nucleic acid assay, adapted to a microsphere array detection platform. We call this assay multiplex oligonucleotide ligation-PCR (MOL-PCR). Unlike other ligation-based assays that require multiple steps, our protocol consists of a single tube reaction, followed by hybridization to a Luminex microsphere array for detection. We demonstrate the ability of this assay to simultaneously detect diverse nucleic acid signatures (e.g., unique sequences, single nucleotide polymorphisms) in a single multiplex reaction. Detection probes consist of modular components that enable target detection, probe amplification, and subsequent capture onto microsphere arrays. To demonstrate the utility of our assay, we applied it to the detection of three biothreat agents, B. anthracis, Y. pestis, and F. tularensis. Combined with the ease and robustness of this assay, the results presented here show a strong potential of our assay for use in diagnostics and surveillance. |
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Authors:
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Alina Deshpande; Jason Gans; Steven W Graves; Lance Green; Laura Taylor; Heung Bok Kim; Yuliya A Kunde; Pascale M Leonard; Po-E Li; Jacob Mark; Jian Song; Momchilo Vuyisich; P Scott White |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S. Date: 2009-12-16 |
Journal Detail:
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Title: Journal of microbiological methods Volume: 80 ISSN: 1872-8359 ISO Abbreviation: J. Microbiol. Methods Publication Date: 2010 Feb |
Date Detail:
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Created Date: 2010-02-01 Completed Date: 2010-03-30 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8306883 Medline TA: J Microbiol Methods Country: Netherlands |
Other Details:
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Languages: eng Pagination: 155-63 Citation Subset: IM |
Copyright Information:
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Published by Elsevier B.V. |
Affiliation:
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Decision Applications Division, Mail Stop K551, Los Alamos National Laboratory, Los Alamos, NM 87545, United States. deshpande_a@lanl.gov |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Bacillus anthracis
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genetics,
isolation & purification Clinical Laboratory Techniques* Francisella tularensis / genetics, isolation & purification Humans Ligase Chain Reaction / methods* Microarray Analysis / methods* Microspheres Molecular Diagnostic Techniques / methods* Yersinia pestis / genetics, isolation & purification |
| Grant Support | |
ID/Acronym/Agency:
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RR001315/RR/NCRR NIH HHS |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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