A new chemically amplified electrochemical system for DNA detection in solution

A newly developed chemically amplified electrochemical detection system was applied to the quantitation of DNA in solution. The system employed Ru(bpy)2dppz (bpy = 2,2′-bipyridine, dppz = dipyrido[3,2-a :2′,3′-c]phenazine), a high-affinity DNA intercalator, as the electrochemical indicator, oxalate as the sacrificial electron donor to chemically amplify the electrochemical signal, and tin-doped indium oxide as the working electrode to suppress background current. Intercalation of Ru(bpy)2dppz into calf thymus DNA in solution led to a reduction in the oxalate-amplified electrochemical current as compared to a DNA-free solution. The degree of reduction was a function of the concentration of DNA, thus forming the basis for DNA detection. To illustrate the advantages of the new system, a direct comparison was made between amplified (with oxalate) and non-amplified (without oxalate) DNA detection. In the presence of 100 mM oxalate, anodic current of Ru(bpy)2dppz was amplified by more than 60 folds, resulting in substantial improvement in signal-to-background ratio. Furthermore, as the DNA concentration was increased, the amplified current decayed much faster than the non-amplified signal, giving rise to higher detection sensitivity. The steeper decay was attributed to slower redox reaction between DNA-intercalated Ru(bpy)2dppz and oxalate, as the negatively charged phosphate groups on DNA repelled the anions. Effect of ionic strength was investigated to provide support for the interpretation. As expected, the decay of the amplified response with increasing concentration of DNA became less steep when more NaCl was added into the solution. The opposite effect was observed when tri-propylamine, a cationic electron donor, was used instead of oxalate. With an optimized concentration of 30 mM oxalate and 5 μM Ru(bpy)2dppz, calf thymus DNA of as low as 1 pM was detected in solution, which was close to the detection limit of some fluorescence measurements.