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Single quantum dot analysis enables multiplexed point mutation detection by gap ligase chain reaction.
- Source :
-
Small (Weinheim an der Bergstrasse, Germany) [Small] 2013 Apr 08; Vol. 9 (7), pp. 1096-105. Date of Electronic Publication: 2012 Dec 13. - Publication Year :
- 2013
-
Abstract
- Gene point mutations present important biomarkers for genetic diseases. However, existing point mutation detection methods suffer from low sensitivity, specificity, and a tedious assay processes. In this report, an assay technology is proposed which combines the outstanding specificity of gap ligase chain reaction (Gap-LCR), the high sensitivity of single-molecule coincidence detection, and the superior optical properties of quantum dots (QDs) for multiplexed detection of point mutations in genomic DNA. Mutant-specific ligation products are generated by Gap-LCR and subsequently captured by QDs to form DNA-QD nanocomplexes that are detected by single-molecule spectroscopy (SMS) through multi-color fluorescence burst coincidence analysis, allowing for multiplexed mutation detection in a separation-free format. The proposed assay is capable of detecting zeptomoles of KRAS codon 12 mutation variants with near 100% specificity. Its high sensitivity allows direct detection of KRAS mutation in crude genomic DNA without PCR pre-amplification.<br /> (Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)
Details
- Language :
- English
- ISSN :
- 1613-6829
- Volume :
- 9
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- Small (Weinheim an der Bergstrasse, Germany)
- Publication Type :
- Academic Journal
- Accession number :
- 23239594
- Full Text :
- https://doi.org/10.1002/smll.201202242