Electrochemical detection of oligonucleotide by attaching redox probes onto its backbone

An approach was demonstrated to detect oligonucleotide by attaching redox probes onto its backbone. First, peptide nucleic acid (PNA) with a neutral backbone was immobilized onto a gold (Au) electrode surface as a capture. Second, when the PNA capture hybridized with a target oligonucleotide (a short DNA), an assembly of Au-PNA-DNA formed and phosphate groups were thus brought into the assembly from the DNA's backbone. The linker ion of Zr(4+) exhibits a strong coordination interaction with the phosphate group and the carboxylic group. The hybridized target DNA provides the phosphate group while a derivatized redox probe of ferrocene (Fc) carboxyl acid offers the carboxylic group. Therefore, the redox probe can be attached to the phosphate group by the linker to form an assembly of Au-PNA-DNA-Zr-Fc. Its redox process was studied and the detection conditions of oligonucleotide were optimized. A limit of detection of 1.0×10(-12) M or ∼2 attomol was reached.