Homogeneous point mutation detection by quantum dot-mediated two-color fluorescence coincidence analysis
This report describes a new genotyping method capable of detecting low-abundant point mutations in a homogeneous, separation-free format. The method is based on integration of oligonucleotide ligation with a semiconductor quantum dot (QD)-mediated two-color fluorescence coincidence detection scheme....
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Published in | Nucleic acids research Vol. 34; no. 5; p. e35 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
England
Oxford University Press
01.01.2006
Oxford Publishing Limited (England) |
Subjects | |
Online Access | Get full text |
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Summary: | This report describes a new genotyping method capable of detecting low-abundant point mutations in a homogeneous, separation-free format. The method is based on integration of oligonucleotide ligation with a semiconductor quantum dot (QD)-mediated two-color fluorescence coincidence detection scheme. Surface-functionalized QDs are used to capture fluorophore-labeled ligation products, forming QD-oligonucleotide nanoassemblies. The presence of such nanoassemblies and thereby the genotype of the sample is determined by detecting the simultaneous emissions of QDs and fluorophores that occurs whenever a single nanoassembly flows through the femtoliter measurement volume of a confocal fluorescence detection system. The ability of this method to detect single events enables analysis of target signals with a multiple-parameter (intensities and count rates of the digitized target signals) approach to enhance assay sensitivity and specificity. We demonstrate that this new method is capable of detecting zeptomoles of targets and achieve an allele discrimination selectivity factor >105. |
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Bibliography: | istex:A75E2DA733382254E899669FB3CAB6007C5D2D22 ark:/67375/HXZ-MV6BVQ18-V To whom correspondence should be addressed. Tel: +1 410 516 7086; Fax: 1 410 516 7254; Email: thwang@jhu.edu local:gkl021 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0305-1048 1362-4962 |
DOI: | 10.1093/nar/gkl021 |