title: Discrimination of single mutations in cancer-related gene fragments with a surface acoustic wave sensor.
abstract: Here, we report on using a surface acoustic wave sensor for the highly sensitive and accurate detection of individual point mutations in cancer-related gene DNA fragments from single injections. Our sensor measures both the mass and viscosity signals and, thus, allows discriminating between mass effects resulting from hybridization of short DNA strands and viscosity effects due to increasing amounts of DNA deposited on the sensor. Single nucleotide exchanges or deletions are distinguished reliably and with exceeding simplicity from the wild-type sequences, on the basis of differences in their dissociation or association rates starting at low nanomolar concentrations. Mutant oligonucleotides were identified immediately from viewing the recorded signal and without further processing of the data. Multiple repeated binding cycles were possible over days without affecting sensitivity. To achieve signal amplification, our new bioassay can also apply multiple hybridization steps based on sandwich hybridizations. Kinetic evaluations gave insight into the physicochemical properties of the fragments used that explain the differences in their binding processes.
authors.affiliation: LIMES Program Unit Chemical Biology & Medicinal Chemistry, c/o Kekulé Institute for Organic Chemistry and Biochemistry, University of Bonn, Gerhard-Domagk-Strasse 1, 53121 Bonn, Germany.
authors.forename: Thomas M A
authors.initials: TM
authors.lastname: Gronewold
authors.forename: Antje
authors.initials: A
authors.lastname: Baumgartner
authors.forename: Eckhard
authors.initials: E
authors.lastname: Quandt
authors.forename: Michael
authors.initials: M
authors.lastname: Famulok
mesh_terms: D000162:Acoustics; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D015342:DNA Probes / Q000235:genetics; D007700:Kinetics; D009154:Mutation / Q000235:genetics; D009363:Neoplasm Proteins / Q000032:analysis* / Q000235:genetics*; D000072079:Patched Receptors; D011956:Receptors, Cell Surface / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000235:genetics
chemical_list: D015342:DNA Probes; D009363:Neoplasm Proteins; D000072079:Patched Receptors; D011956:Receptors, Cell Surface; D016159:Tumor Suppressor Protein p53