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基于外切核酸酶 III 的荧光适体传感器用于检测癌胚抗原:石墨烯氧化物和 2-氨基嘌呤的应用。

Exonuclease III-assisted fluorometric aptasensor for the carcinoembryonic antigen using graphene oxide and 2-aminopurine.

机构信息

School of Life Sciences, Central South University, Changsha, 410013, China.

出版信息

Mikrochim Acta. 2019 Jul 3;186(8):500. doi: 10.1007/s00604-019-3621-4.

Abstract

A reliable fluorometric assay is described for the determination carcinoembryonic antigen (CEA) using exonuclease III (Exo III) and a 2-aminopurine binding aptamer. In the absence of CEA, dsDNA is degraded by Exo III, and free 2-AP (which has a blue fluorescence with excitation/emission maxima of 310/365 nm) is released. Strong fluorescence is generated after addition of graphene oxide (GO) to the solution. However, the 2-AP modified DNA (T2) cannot be degraded in the presence of CEA by Exo III due to the interaction between CEA and aptamer T1. Hence, only weak fluorescence can be detected after addition of GO. In this system, CEA can be quantified in the 0.05 - 2 ng·mL concentration range with a detection limit of 30 pg·mL (at S/N = 3). The method was successfully applied to analyze serum samples for CEA. Graphical Abstract An exonuclease III-assisted fluorometric aptasensor has been developed for the detection of carcinoembryonic antigen using graphene oxide and 2-aminopurine.

摘要

一种可靠的荧光分析方法被描述用于使用exonuclease III(Exo III)和 2-氨基嘌呤结合适体测定癌胚抗原(CEA)。在不存在 CEA 的情况下,dsDNA 被 Exo III 降解,游离的 2-AP(其荧光激发/发射最大值为 310/365nm)被释放。向溶液中加入氧化石墨烯(GO)后,会产生强烈的荧光。然而,由于 CEA 和适体 T1 之间的相互作用,在存在 CEA 的情况下,Exo III 不能使 2-AP 修饰的 DNA(T2)降解。因此,加入 GO 后只能检测到弱荧光。在该体系中,CEA 可在 0.05-2ng·mL 的浓度范围内定量,检测限为 30pg·mL(S/N=3)。该方法成功地应用于血清样品中 CEA 的分析。 图摘要 一种基于氧化石墨烯和 2-氨基嘌呤的 exonuclease III 辅助荧光适体传感器已被开发用于检测癌胚抗原。

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