An electrochemical aptasensor for the highly sensitive detection of 8-hydroxy-2'-deoxyguanosine based on the hybridization chain reaction

Abstract

In the present work a highly sensitive and selective aptasensor was developed for the determination of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) based on the hybridization chain reaction (HCR) signal amplification. It was observed that the aptamer of 8-OH-dG could hybridize with the capture DNA immobilized on the gold electrode with a sticky tail left, which initiated the HCR and led to the formation of extended dsDNA structure on the electrode surface. Then the electroactive species ([Ru(NH₃)₆]³⁺, RuHex) intercalated into the dsDNA grooves to generate the amplified signal. However, in the presence of 8-OH-dG, the aptamer containing G-rich nucleic acid sequences would be induced to form a G-quadruplex structure, which made it impossible to continue the HCR. So the detection signal will significantly decrease. Under the optimal conditions, the peak current of RuHex was linear with the logarithm of 8-OH-dG concentration in the range from 10pM to 100μM with the detection limit of 2.5pM. By integrating the merits of enzyme-free amplification power of the HCR and the inherent high sensitivity of the electrochemical technique, the prepared aptasensor not only showed high sensitivity for the detection of 8-OH-dG, but also exhibited good selectivity against to the uric acid, an important interferent in the urine sample. Particularly, the aptasensor was applied to detect 8-OH-dG in urine samples with satisfactory results.

Keywords: 8-Hydroxy-2′-deoxyguanosine; Electrochemical aptasensor; G-quadruplex; Hybridization chain reaction.