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Phthalic Acid Ester-Binding DNA Aptamer Selection, Characterization, and Application to an Electrochemical Aptasensor
Published on: March 21, 2018
Acetamiprid multidetection by disposable electrochemical DNA aptasensor
R Rapini1, A Cincinelli1, G Marrazza1
1Department of Chemistry "Ugo Schiff", University of Florence, Via della Lastruccia 3, Sesto Fiorentino (FI) 50019, Italy.
Abstract:
In this work, we propose an electrochemical DNA aptasensor for sensitive multidetection of acetamiprid based on a competitive format and disposable screen-printed arrays. To improve the sensitivity of the aptasensor, polyaniline film and gold nanoparticles were progressively electrodeposited on the graphite screen-printed electrode surface by cyclic voltammetry. Gold nanoparticles were then employed as platform for thiol-tethered DNA aptamer immobilization. Different acetamiprid solutions containing a fixed amount of biotinylated complementary oligonucleotide sequence by DNA aptasensor arrays were analyzed. Streptavidin-alkaline phosphatase conjugate was then added to trace the affinity reaction. The enzyme catalyzed the hydrolysis of 1-naphthyl phosphate to 1-naphthol. The enzymatic product was detected by differential pulse voltammetry. A decrease of the signal was obtained when the pesticide concentration was increased, making the sensor work as signal off sensor. Under optimized conditions by testing key experimental parameters, a dose-response curve was constructed between 0.25 and 2.0µM acetamiprid concentration range and a limit of detection of 0.086µM was calculated. The selectivity of the aptasensor was also confirmed by the analysis of atrazine pesticide. Finally, preliminary experiments in fruit juice samples spiked with acetamiprid were also performed.
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