Selective determination of non-organophosphorus insecticide using DNA aptamer-based single-use biosensors

In the present study, we developed a disposable aptamer-based biosensor for rapid, sensitive, and reliable detection of acetamiprid (ACE). To improve the sensitivity of the aptasensor, poly-5-amino-2-mercapto-1,3,4-thiadiazole [P(AMT)] and gold nanoparticles (AuNPs) were progressively electrodeposited on the screen-printed electrode (SPE) surface by using cyclic voltammetry (CV) technique. For the determination of ACE, thiol-modified primary aptamer (Apt1) was selected by using the SELEX method and immobilized on the surface of the P(AMT) and AuNPs-modified SPE (SPE/P(AMT)/AuNPs) via Au-S bonding. Then, the surface-bound aptamer was incubated with ACE for 45 Min. After that, the biotin-labeled aptamer 2 (Apt2) was interacted with the ACE, then the enzyme-labeled step was performed. In this step, alkaline phosphatase (ALP) was bound to the surface through the interaction between Apt2 labeled with biotin and streptavidin (strep)-ALP conjugate. The determination of ACE was achieved by measuring the oxidation signal of alpha-naphthol, which is formed on the electrode surface through the interaction of ALP with alpha-naphthyl phosphate. The working range of the developed aptasensor was determined as 5 x 10(-12)-5 x 10(-10 )mol L(-1)with a low limit of detection (1.5 pmol L-1). It was also found that the proposed aptasensor possessed great advantages such as low cost, good selectivity, and good reproducibility.