Redox-active nucleic-acid replica for the amplified bioelectrocatalytic detection of viral DNA.

A new concept for the amplified electrochemical detection of the 7229-base viral DNA of M13phi is developed. A thiolated 27-base nucleic acid (1) is assembled on an Au-electrode. Hybridization between the sensing interface and the M13phi DNA is followed by the polymerase-induced replication of the analyte DNA in the presence of dCTP, dGTP, dATP, and ferrocene-tethered-dUTP (2). The generated redox-active replica mediates electron transfer between the enzyme glucose oxidase (GOx) and the electrode and activates the bioelectrocatalyzed oxidation of glucose. The bioelectrocatalyzed oxidation of glucose provides a biocatalytic amplification path for the formation of the redox-active replica. The electrochemical techniques to follow the replication and the bioelectrocatalytic amplification are differential pulse voltammetry and cyclic voltammetry. The electrical responses from the system relate to the bulk concentration of the M13phi DNA, thus enabling the quantitative analysis of the viral gene.