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A rapid multiplex assay for nucleic acid-based diagnostics.
- Source :
-
Journal of microbiological methods [J Microbiol Methods] 2010 Feb; Vol. 80 (2), pp. 155-63. Date of Electronic Publication: 2009 Dec 16. - Publication Year :
- 2010
-
Abstract
- 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.<br /> (Published by Elsevier B.V.)
- Subjects :
- Bacillus anthracis genetics
Bacillus anthracis isolation & purification
Francisella tularensis genetics
Francisella tularensis isolation & purification
Humans
Microspheres
Yersinia pestis genetics
Yersinia pestis isolation & purification
Clinical Laboratory Techniques
Ligase Chain Reaction methods
Microarray Analysis methods
Molecular Diagnostic Techniques methods
Subjects
Details
- Language :
- English
- ISSN :
- 1872-8359
- Volume :
- 80
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Journal of microbiological methods
- Publication Type :
- Academic Journal
- Accession number :
- 20006656
- Full Text :
- https://doi.org/10.1016/j.mimet.2009.12.001