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利用钆离子作为亲和探针,从复杂样品中选择性地富集和磁分离细菌。

Using gadolinium ions as affinity probes to selectively enrich and magnetically isolate bacteria from complex samples.

机构信息

Department of Applied Chemistry, National Chiao Tung University, Hsinchu, 300, Taiwan.

Department of Applied Chemistry, National Chiao Tung University, Hsinchu, 300, Taiwan.

出版信息

Anal Chim Acta. 2020 May 29;1113:18-25. doi: 10.1016/j.aca.2020.03.046. Epub 2020 Mar 28.

Abstract

Magnetic trapping has been employed in the development of analytical methods owing to its ease and simplicity in handling samples. Nevertheless, the generation of functional probes is usually time consuming. A new and simple affinity method that uses gadolinium ion (Gd), a magnetic ion, as affinity probe for magnetic tapping of pathogenic bacteria was demonstrated in the present study. Escherichia coli O157:H7, Staphylococcus aureus, and Acinetobacter baumannii were selected as model bacteria. The model bacteria were magnetically isolated after incubation in Tris buffer (pH 8) containing Gd (0.1 M) under microwave heating (power: 180 W, 90 s × 3). The resultant Gd-bacterium conjugates possessed sufficient magnetism, resulting in magnetic aggregations by an external magnet (∼4,000 Gauss). For ease of magnetic isolation, the sample containing Gd-bacterium complexes was stirred by a small magnet. After 1 h, the magnet attached with precipitates, i.e., Gd-bacterium conjugates, was readily removed using a pair of tweezers. The bacteria in the resultant conjugates were characterized by matrix-assisted laser desorption/ionization mass spectrometry. The limits of detection of the current approach toward E. coli O157:H7, S. aureus, and A. baumannii in complex samples were ∼10-10 cells mL.

摘要

由于操作简便,磁捕获技术已被应用于分析方法的开发中。然而,功能探针的生成通常耗时较长。本研究提出了一种新的简单亲和方法,使用顺磁离子钆(Gd)作为亲和探针,用于磁性捕获致病菌。选择大肠杆菌 O157:H7、金黄色葡萄球菌和鲍曼不动杆菌作为模型菌。在含有 Gd(0.1 M)的 Tris 缓冲液(pH 8)中孵育后,模型菌在微波加热(功率:180 W,90 s×3)下被磁性分离。所得 Gd-细菌缀合物具有足够的磁性,在外磁场(约 4000 高斯)下产生磁性聚集。为了便于磁性分离,用小磁铁搅拌含有 Gd-细菌复合物的样品。1 小时后,用一对镊子很容易地将附有沉淀物(即 Gd-细菌缀合物)的磁铁取出。用基质辅助激光解吸/电离质谱法对所得缀合物中的细菌进行了表征。该方法对复杂样品中大肠杆菌 O157:H7、金黄色葡萄球菌和鲍曼不动杆菌的检测限约为 10-10 个细胞/mL。

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