title: A label-free, electrochemical SERS-based assay for detection of DNA hybridization and discrimination of mutations.
abstract: A label-free, surface-enhanced Raman spectroscopy-based assay for detecting DNA hybridization at an electrode surface and for distinguishing between mutations in DNA is demonstrated. Surface-immobilized DNA is exposed to a binding agent that is selective for dsDNA and acts as a reporter molecule. Upon application of a negative potential, the dsDNA denatures into its constituent strands, and the changes in the spectra of the reporter molecule are monitored. This method has been used to distinguish between a wild-type, 1653C/T single-point mutation and ΔF508 triplet deletion in the CFTR gene. The use of dsDNA-selective binding agents as reporter molecules in a discrimination assay removes the burden of synthetically modifying the target to be detected, while retaining flexibility in the choice of the reporter molecule.
authors: School of Chemistry, University of Southampton, Southampton SO17 1BJ, United Kingdom.
authors: Robert P
authors: RP
authors: Johnson
authors: James A
authors: JA
authors: Richardson
authors: Tom
authors: T
authors: Brown
authors: Philip N
authors: PN
authors: Bartlett
mesh_terms: D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods; D004396:Coloring Agents / Q000737:chemistry; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D061925:Immobilized Nucleic Acids / Q000737:chemistry; D007364:Intercalating Agents / Q000737:chemistry; D009693:Nucleic Acid Hybridization / Q000379:methods*; D017354:Point Mutation; D013059:Spectrum Analysis, Raman / Q000379:methods*
chemical_list: D004396:Coloring Agents; D061925:Immobilized Nucleic Acids; D007364:Intercalating Agents; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA