Photoelectrochemical DNA sensor for the rapid detection of DNA damage induced by styrene oxide and the Fenton reaction

Rapid and sensitive detection methods are in urgent demand for the screening of an overwhelming number of existing and new chemicals as potential DNA-damaging agents. In this study, two photoelectrochemistry-based DNA sensor configurations were employed in the detection of DNA damage caused by styrene oxide and Fe2+/H2O2. The organic compound and heavy metal represent genotoxic chemicals possessing two major damaging mechanisms, DNA adduct formation and DNA oxidation. In the first sensor configuration, a ruthenium tris(bipyridine)-labeled avidin film and a double-stranded calf thymus DNA (ds-DNA) film were assembled successively on tin oxide nanoparticle film electrodes. Photogenerated Ru(III) oxidized guanidine and adenosine bases in DNA and gave rise to photocurrent. DNA damage was detected after the reaction of the DNA film with either styrene oxide or Fe2+/H2O2, which exposed more DNA bases for photooxidation and resulted in increased photocurrent. In the second configuration, an unlabeled avidin film and a ds-DNA film were assembled on the semiconductor electrode. A DNA intercalator, Ru(bpy)(2)(dppz)(2+) (bpy = 2,2'-bipyridine, dppz = dipyrido[3,2-a:2',3'-c]phenazine), was employed as the photoelectrochemical signal reporter. After the chemical reaction with the damaging agents, the DNA film bound less Ru(bpy)(2)(dppz)(2+), accompanied by a drop in photocurrent. Both sensors were used to follow the reaction course in styrene oxide and Fenton reagents and produced similar results. According to the data, damage of the DNA film was complete in 1 h in Fenton reagents and in 3 h in styrene oxide. In addition, the Fenton reaction induced much more severe damage than styrene oxide. The results demonstrate for the first time that the photoelectrochemical DNA sensor can detect both DNA adduct formation and DNA oxidation. It has the potential of becoming a screening tool for the rapid assessment of the genotoxicity of existing and new chemicals.