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Selective determination of non-organophosphorus insecticide using DNA aptamer-based single-use biosensors
Biotechnology and Applied Biochemistry ( IF 3.2 ) Pub Date : 2020-09-24 , DOI: 10.1002/bab.2039
Mustafa Barış Koçer 1 , Gözde Aydoğdu Tığ 2 , Şule Pekyardımcı 2
Affiliation  

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 AuS 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 α-naphthol, which is formed on the electrode surface through the interaction of ALP with α-naphthyl phosphate. The working range of the developed aptasensor was determined as 5 × 10−12–5 × 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.

中文翻译:

使用基于 DNA 适配体的一次性生物传感器选择性测定非有机磷杀虫剂

在本研究中,我们开发了一种基于适配体的一次性生物传感器,用于快速、灵敏、可靠地检测啶虫脒 (ACE)。为了提高适体传感器的灵敏度,将聚 5-氨基-2-巯基-1,3,4-噻二唑 [P(AMT)] 和金纳米粒子 (AuNPs) 逐步电沉积在丝网印刷电极 (SPE) 表面上通过使用循环伏安法(CV)技术。为了测定 ACE,采用 SELEX 方法选择硫醇修饰的初级适体 (Apt1),并通过 Au 固定在 P(AMT) 和 AuNPs 修饰的 SPE (SPE/P(AMT)/AuNPs) 的表面上S 键合。然后,将表面结合的适体与 ACE 一起孵育 45 分钟。之后,生物素标记的适体2(Apt2)与ACE相互作用,然后进行酶标记步骤。在此步骤中,碱性磷酸酶 (ALP) 通过生物素标记的 Apt2 与链霉亲和素 (strep)-ALP 偶联物之间的相互作用与表面结合。ACE的测定是通过测量α-萘酚的氧化信号来实现的,α-萘酚是通过ALP与α-萘基磷酸盐的相互作用在电极表面形成的。开发的适体传感器的工作范围被确定为 5 × 10 -12 –5 × 10 -10  mol L -1具有低检测限(1.5 pmol L -1)。还发现所提出的适体传感器具有成本低、选择性好、重现性好等优点。
更新日期:2020-09-24
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