Sequence-specific analysis of oligodeoxynucleotides by precipitate-amplified surface plasmon resonance measurements

Dongming Zeng
2007 Frontiers in Bioscience  
A flow injection SPR spectrometer is used to detect DNA hybridization via amplification by an enzymecatalyzed precipitation reaction. Hybridization of an oligodeoxynucleotide (ODN) target with a surfaceconfined ODN probe, followed by hybridization between the overhang on the resultant duplex and a biotin-tagged ODN, yields a "sandwich" complex (a three-component double-stranded). A streptavidin-horseradish peroxidase conjugate (SA-HRP) can be attached to the sandwich complex-covered surface via
more » ... the biotin/streptavidin complexation. In the presence of H 2 O 2 , the HRP catalyzes oxidation of 4-chloro-1-naphthol (CN) to form a precipitate on the sensor surface. The precipitated film dramatically changes the refractive index at the metal/dielectric interface and significantly lowers the detection level. The method is shown to be reproducible, to possess high ODN sequence specificity and a high sensitivity allowing detection of ODN target concentration as low as 10 fM.
doi:10.2741/2552 pmid:17569634 fatcat:tyh65sysfbcjza3h4q24vzfeq4