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使用适配体功能化磁性二氧化硅固相和MCM - 41 - 适配体门控系统对牛奶样品进行快速灵敏检测

Fast and Sensitive Detection of in Milk Samples Using Aptamer-Functionalized Magnetic Silica Solid Phase and MCM-41-Aptamer Gate System.

作者信息

Bayramoglu Gulay, Ozalp V Cengiz, Dincbal Uguray, Arica M Yakup

机构信息

Biochemical Processing and Biomaterial Research Laboratory, Gazi University, 06500 Teknikokullar, Ankara, Turkey.

Department of Chemistry, Faculty of Sciences, Gazi University, 06500 Ankara, Turkey.

出版信息

ACS Biomater Sci Eng. 2018 Apr 9;4(4):1437-1444. doi: 10.1021/acsbiomaterials.8b00018. Epub 2018 Mar 12.

Abstract

General detection methods for include PCR analysis, immunologic methods, solid culturing techniques, and various microscopic studies. Milk and other food samples demonstrate an especially difficult challenge for direct detection, resulting from high biological contents. In this report, we aimed for fast detection of pathogen cells through an efficient magnetic capture and subsequent quick detection based on aptamer affinity. The FeO@SiO@pGMA and MCM-41 particles were prepared separately and used for preconcentration and detection, respectively. Aptamer oligonucleotide sequences against were fixed on both amine-functionalized MCM-41 and FeO@SiO@pGMA particles via glutaraldehyde coupling. The captured cells were determined by a fluorescent homogeneous assay in the samples by aptamer-gated MCM-41 silica particles. Our method achieved a sensitive assay to detect cells in milk samples as low as 10 CFU/ml without any culturing. Hence, the proposed sensing strategy might be an efficient platform for pathogen detection in a food matrix.

摘要

通用的检测方法包括聚合酶链反应(PCR)分析、免疫方法、固体培养技术以及各种显微镜研究。由于生物成分含量高,牛奶和其他食品样本的直接检测面临特别严峻的挑战。在本报告中,我们旨在通过高效的磁性捕获以及随后基于适配体亲和力的快速检测来实现对病原体细胞的快速检测。分别制备了FeO@SiO@pGMA和MCM - 41颗粒,并分别用于预浓缩和检测。通过戊二醛偶联,将针对[病原体名称未给出]的适配体寡核苷酸序列固定在胺功能化的MCM - 41和FeO@SiO@pGMA颗粒上。通过适配体门控的MCM - 41二氧化硅颗粒,采用荧光均相分析法测定样品中捕获的[病原体名称未给出]细胞。我们的方法实现了对牛奶样本中低至10 CFU/ml的[病原体名称未给出]细胞进行灵敏检测,无需任何培养。因此,所提出的传感策略可能是用于食品基质中病原体检测的有效平台。

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