title: SERS primers and their mode of action for pathogen DNA detection.
abstract: SERS primers have been used to directly detect specific PCR products utilizing the difference in adsorption of single-stranded and double-stranded DNA onto nanoparticle surfaces. Herein, seven parameters important for improved positive SERS assays for real applications were investigated. First, we applied a model system for optimization experiments, followed by a PCR assay to detect pathogen DNA, and then the introduction of a new assay that utilizes the 5'→3' exonuclease activity of Taq DNA polymerase to partly digest the SERS probe, generating dye-labeled single-stranded DNA increasing the SERS signals for detection of pathogen DNA. Applying the model system, it was found that uni-molecular SERS primers perform better than bi-molecular SERS primers. However, within the PCR assays, it was found that uni- and bi-molecular SERS primers performed very similarly, and the most reproducible results were obtained using the 5'→3' exonuclease digestion assay. These SERS-based assays offer new routes over conventional fluorescence-based techniques without compromising sensitivity or selectivity.
authors: Centre for Molecular Nanometrology, WestCHEM, Pure and Applied Chemistry, University of Strathclyde, 295 Cathedral Street, Glasgow G1 1XL, United Kingdom.
authors: Danny
authors: D
authors: van Lierop
authors: Iain A
authors: IA
authors: Larmour
authors: Karen
authors: K
authors: Faulds
authors: Duncan
authors: D
authors: Graham
mesh_terms: D017931:DNA Primers / Q000032:analysis* / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D013059:Spectrum Analysis, Raman / Q000379:methods*
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial