Label-Free Detection of C–T Mutations by Surface-Enhanced Raman Spectroscopy Using Thiosulfate-Modified Nanoparticles

Recent developments in molecular spectroscopy have widened the scope of surface-enhanced Raman spectroscopy (SERS) for detection of nucleic acids. In order to solve the interference of impurity signals in SERS analysis that hamper the reliable detection of DNA, Ag nanoparticles modified with thiosulfate ions were used to obtain SERS signals of DNA molecules in aqueous solutions, which showed good reproducibility. By using thiosulfate ions and calcium ions as aggregating agents, this method not only eliminated the influence of citrate on DNA signals completely but also obtained the signals for all bases indiscriminately, including the T base that was considered to have low Raman activity. Subsequently, the base stacking rule was used to identify mutations arising from C/T transition. It further identified the mutation sites of single-base C/T transition using this platform for the first time. This method has wide application prospects in DNA analysis, DNA sequencing, and genetic testing.