Colloidal gold–polystyrene bead hybrid for chemiluminescent detection of sequence-specific DNA

Abstract

A sensitive chemiluminescent (CL) detection of sequence-specific DNA has been developed by taking advantage of a magnetic separation/mixing process and the amplification feature of Colloidal gold labels. In this protocol, the target oligonucleotides are hybridized with magnetic bead-linked capture probes, followed by the hybridization of the biotin-terminated amplifying DNA probes and the binding of streptavidin-coated gold nanoparticles; the nanometer-sized gold tags are then dissolved and quantified by a simple and sensitive luminol CL reaction. The proposed CL protocol is evaluated for a 30-base model DNAsequence, and the amount as low as 0.01 pmol of DNA is determined, which exhibits a 150× enhancement in sensitivity over previous gold dissolution-based electrochemical formats and an enhancement of 20× over the ICPMS detection. Further signal amplification is achieved by the assembly of biotinylated Colloidal gold onto the surface of streptavidin-coated polystyrene beads. Such amplified CL transduction allows detection of DNA targets down to the 100 amol level, and offers great promise for ultrasensitive detection of other biorecognition events.