DNA‐probes for the highly sensitive identification of single nucleotide polymorphism using single‐molecule spectroscopy
Authors:
- Nicole MarméNMNicole Marmé
- Jens-Peter KnemeyerJKJens-Peter Knemeyer
- 20 March 2007
- journal article
- Published by Wiley in FEBS Letters
- Vol. 581 (8) , 1644-1648
Abstract
This article presents a new, highly sensitive method for the identification of single nucleotide polymorphisms (SNPs) in homogeneous solutions using fluorescently labeled hairpin‐structured oligonucleotides (smart probes) and fluorescence single‐molecule spectroscopy. While the hairpin probe is closed, fluorescence intensity is quenched due to close contact between the chromophore and several guanosine residues. Upon hybridization to the respective target SNP sequence, contact is lost and the fluorescence intensity increases significantly. High specificity is achieved by blocking sequences containing mismatch with unlabeled oligonucleotides. Time‐resolved single‐molecule fluorescence spectroscopy enables the detection of individual smart probes passing a small detection volume. This method leads to a subnanomolar sensitivity for this single nucleotide specific DNA assay technique.Keywords
This publication has 20 references indexed in Scilit:
- Identification of single-point mutations in mycobacterial 16S rRNA sequences by confocal single-molecule fluorescence spectroscopyNucleic Acids Research, 2006
- Sequencing-Based Detection of Low-Frequency Human Immunodeficiency Virus Type 1 Drug-Resistant Mutants by an RNA/DNA Heteroduplex Generator-Tracking AssayJournal of Virology, 2004
- Direct Detection of Rifampin- and Isoniazid-Resistant Mycobacterium tuberculosis in Auramine-Rhodamine-Positive Sputum Specimens by Real-Time PCRJournal of Clinical Microbiology, 2004
- Development and Validation of a Quantitative Real-Time PCR Assay Using Fluorescence Resonance Energy Transfer Technology for Detection of Aspergillus fumigatus in Experimental Invasive Pulmonary AspergillosisJournal of Clinical Microbiology, 2003
- Highly sensitive detection of the MGB1 transcript (mammaglobin) in the peripheral blood of breast cancer patientsInternational Journal of Cancer, 2003
- Photoinduced Electron Transfer between Fluorescent Dyes and Guanosine Residues in DNA-HairpinsThe Journal of Physical Chemistry B, 2003
- New Real-Time PCR Able To Detect in a Single Tube Multiple Rifampin Resistance Mutations and High-Level Isoniazid Resistance Mutations in Mycobacterium tuberculosisJournal of Clinical Microbiology, 2002
- Multiplex detection of single-nucleotide variations using molecular beaconsGenetic Analysis: Biomolecular Engineering, 1999
- Detection and identification of single dye labeled mononucleotide molecules released from an optical fiber in a microcapillary: First steps towards a new single molecule DNA sequencing techniquePhysical Chemistry Chemical Physics, 1999
- Molecular Beacons: Probes that Fluoresce upon HybridizationNature Biotechnology, 1996