Electrical Detection of Oligonucleotide Using an Aggregate of Gold Nanoparticles as a Conductive Tag

Abstract

Sequence-specific DNA detection is a routine job in medical diagnostics and genetic screening. Alternative to a fluorescence readout scheme or electrophoresis approach, various kinds of rapid, low-cost, facile, and label-free methods have also been developed in last decades. Among these, direct electrical detection of DNA received increasing attention but more research is desirable. Particularly, enhancement with high discrimination must be employed to selectively amplify the responding signal. A chip-based biosensor was developed in this work to electrically detect 22-mer oligonucleotide DNA at low concentration, from 50 fM to 10 pM. First, a gold nanoparticle (NP) was capped with 3-mercaptopropionic acid through a thiol-gold bond. The derivatized carboxylic acid group showed strong complex interaction with an inorganic linker, Zr(4+). As a result, Zr(4+) could link several hundreds of individual gold NPs together to form an aggregate of nanoparticles (ANP), which was capable of being used as a conductive tag for the electrical detection of DNA. Second, in order to achieve the discriminative localization of ANP to bridge two comb-shaped electrodes (with height of approximately 50 nm and interdistance of 300-350 nm) gapped with insulative material of silicon oxide, peptide nucleic acids were covalently bonded to the silicon oxide in the gap as capture sites for DNA. After hybridization with target DNA, the charged phosphate-containing backbone of DNA was introduced into the gap. Phosphate groups also exhibited strong complex interaction with the linker of Zr(4+) and could react with the residual Zr(4+) on the ANP surface. As a consequence, the conductive tags were linked to the phosphate groups and localized into the gap, which could modify the conductance between the two comb-shaped electrodes in turn. The degree of modification correlated directly to the amount of hybridized DNA and to the concentration of target DNA in sample solution. Compared with the individual NPs used as the tag, a strong enhancement from the gold ANP was obtained.