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基于酶包被脂质体的高灵敏比色适体传感器用于赭曲霉毒素 A 的检测。

Highly sensitive colorimetric aptasensor for ochratoxin A detection based on enzyme-encapsulated liposome.

机构信息

MOE Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China.

MOE Key Laboratory for Analytical Science of Food Safety and Biology, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, College of Chemistry, Fuzhou University, Fuzhou, Fujian 350116, China; College of Biological Science and Engineering, Fuzhou University, Fuzhou, Fujian 350116, China.

出版信息

Anal Chim Acta. 2018 Mar 9;1002:90-96. doi: 10.1016/j.aca.2017.11.061. Epub 2017 Nov 27.

Abstract

A simple, low-cost, and sensitive liposome-based colorimetric aptasensor has been developed to detect ochratoxin A (OTA). Specifically, a dumbbell-shaped probe was designed, including magnetic beads (MBs), double-stranded DNA (dsDNA), and enzyme-encapsulated liposome. The dsDNA formed by the hybridization between OTA aptamer and its complementary probe. And the dsDNA was used to contact the MBs and the enzyme-encapsulated liposome. In the presence of OTA, the aptamer preferred to combine with OTA to form G-quadruplex, resulting in the release of the detection probe and the enzyme-encapsulated liposome. Each liposome contained a large amount of HRP. Thus, when the liposome was lysed by adding the mixed solution of 3,3',5,5'-tetramethylbenzidine (TMB) and HO, a large number of HRP were released. HRP could catalyze the HO-mediated oxidation of TMB and hence resulted in the color change from colorless to blue with the OTA concentration varying, and this variation can be observed by naked eyes easily. The result showed that the absorption intensity at 652 nm enhanced with the increase of OTA concentration ranging from 0.05 to 2.0 ng mL, and the limit of detection was calculated to be 0.023 ng mL (S/N = 3). The developed colorimetric aptasensor has been applied to detect OTA concentration in corn samples with satisfied results.

摘要

一种简单、低成本、灵敏的基于脂质体的比色适体传感器已被开发用于检测赭曲霉毒素 A(OTA)。具体来说,设计了一种哑铃形探针,包括磁性珠(MBs)、双链 DNA(dsDNA)和包被酶的脂质体。dsDNA 是由 OTA 适体与其互补探针杂交形成的。dsDNA 用于与 MBs 和包被酶的脂质体接触。在 OTA 存在下,适体优先与 OTA 结合形成 G-四链体,导致检测探针和包被酶的脂质体释放。每个脂质体都含有大量的 HRP。因此,当通过加入 3,3',5,5'-四甲基联苯胺(TMB)和 HO 的混合溶液裂解脂质体时,会释放出大量的 HRP。HRP 可以催化 HO 介导的 TMB 氧化,从而导致颜色从无色变为蓝色,随着 OTA 浓度的变化,这种变化可以用肉眼轻松观察到。结果表明,在 0.05 至 2.0 ng mL 的 OTA 浓度范围内,652 nm 处的吸收强度随着 OTA 浓度的增加而增强,检测限计算为 0.023 ng mL(S/N = 3)。所开发的比色适体传感器已应用于检测玉米样品中的 OTA 浓度,结果令人满意。

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