Ultrasensitive electrochemical aptasensing of kanamycin antibiotic by enzymatic signal amplification with a horseradish peroxidase-functionalized gold nanoprobe

Abstract Based on the signal transduction with a horseradish peroxidase (HRP)-functionalized gold nanoprobe, this work develops a new electrochemical aptasensing method for kanamycin (Kana) detection. The aptasensor was constructed through the hybridization of biotinylated Kana-aptamer at its complementary oligonucleotide strand-modified electrode, followed by intercalating methylene blue (MB) into the formed double-stranded DNA (dsDNA). Then, the streptavidin and high-content HRP functionalized gold nanoparticle probes were bound onto the aptasensor. As the MB-mediated HRP-catalytic reaction could produce sensitive electrochemical signal and the aptamer-biorecognition toward Kana could cause quantitative decrease of MB intercalation and nanoprobe capture, convenient electrochemical signal transduction was achieved. Both the enzymatic reaction and nanoprobe signal amplification greatly enhance the electrochemical signal, offering ultrahigh sensitivity of the method. The MB intercalation into dsDNA not only provides necessary electron mediator for the enzymatic reaction but also simplifies the electrochemical measurement. Under optimal conditions, this method showed a wide linear range over four-order of magnitude with a low detection limit of 0.88 pg/mL. In addition, the aptasensor features high specificity, excellent repeatability and stability as well as satisfactory reliability. Thus it possesses valuable application potentials.