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用于孔雀石绿荧光生物传感检测的单链DNA适配体的筛选与应用

Selection and Application of ssDNA Aptamers for Fluorescence Biosensing Detection of Malachite Green.

作者信息

Xie Miaojia, Chen Zanlin, Zhao Fengguang, Lin Ying, Zheng Suiping, Han Shuangyan

机构信息

Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological Engineering, South China University of Technology, Guangzhou 510006, China.

School of Light Industry and Engineering, South China University of Technology, Guangzhou 510006, China.

出版信息

Foods. 2022 Mar 10;11(6):801. doi: 10.3390/foods11060801.

Abstract

Malachite green oxalate (MG) is a kind of veterinary drug, which is freely soluble in water and hazardous to aquatic products, resulting in food toxicity and human health problems. The demand for effective and sensitive detection of MG residues is increasing in food safety. In this work, three DNA aptamers MG-36-12/16/17 targeting MG with good affinity (K values were 169.78, 71.94, and 102.46 μM, respectively) were obtained by Capture-SELEX. Furthermore, MG-36-12, MG-76-16-6A, and MG-36-17 were found to perform sensitively and specifically to detect MG as a sensing probe in a FRET fluorescent aptasensor, where the FAM-labeled aptamer and GO were employed as efficient energy donor–acceptor pair. The linear range of this aptasensor using aptamer MG-36-12 was from 1.71 to 514.29 ng/mL and the LOD was as low as 0.79 ng/mL. Additionally, the fluorescent assay using aptamer MG-36-17 to detect MG exhibited a linear relationship from 1.71 to 857.14 ng/mL and a LOD of 2.13 ng/mL. Meanwhile, the aptasensor showed high specificity to MG with no cross-reactivity to other veterinary drugs and had a mean recovery of 81.54% to 100.96% in actual water samples from the aquatic product market.

摘要

草酸孔雀石绿(MG)是一种兽药,可自由溶于水,对水产品有害,会导致食品毒性和人类健康问题。在食品安全领域,对有效且灵敏地检测MG残留的需求日益增加。在这项工作中,通过Capture-SELEX获得了三种对MG具有良好亲和力的DNA适配体MG-36-12/16/17(K值分别为169.78、71.94和102.46 μM)。此外,发现MG-36-12、MG-76-16-6A和MG-36-17在FRET荧光适配体传感器中作为传感探针能灵敏且特异地检测MG,其中FAM标记的适配体和氧化石墨烯(GO)被用作高效的能量供体-受体对。使用适配体MG-36-12的这种适配体传感器的线性范围为1.71至514.29 ng/mL,检测限低至0.79 ng/mL。此外,使用适配体MG-36-17检测MG的荧光测定法显示线性关系为1.71至857.14 ng/mL,检测限为2.13 ng/mL。同时,该适配体传感器对MG表现出高特异性,对其他兽药无交叉反应,在水产品市场的实际水样中的平均回收率为81.54%至100.96%。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/78ed/8953565/29acca1c58d3/foods-11-00801-sch001.jpg

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