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细菌与离子交换颗粒的相互作用及其在基质辅助激光解吸/电离质谱法分离水中细菌鉴定中的潜在应用。

Interaction of bacteria and ion-exchange particles and its potential in separation for matrix-assisted laser desorption/ionization mass spectrometric identification of bacteria in water.

机构信息

Centre for Advanced Water Technology, Public Utilities Board, 82 Toh Guan Road East, WaterHub, #C4-03, Singapore 608576, Singapore.

出版信息

Rapid Commun Mass Spectrom. 2009 Dec;23(24):3983-93. doi: 10.1002/rcm.4338.

Abstract

Identification of microbial contaminants in drinking water is a challenge to matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) due to low levels of microorganisms in fresh water. To avoid the time-consuming culture step of obtaining enough microbial cells for subsequent MALDI-MS analysis, a combination of membrane filtration and nanoparticles- or microparticles-based magnetic separation is a fast and efficient approach. In this work, the interaction of bacteria and fluidMAG-PAA, a cation-exchange superparamagnetic nanomaterial, was investigated by MALDI-MS analysis and transmission electron microscopy. FluidMAG-PAA selectively captured cells of Salmonella, Bacillus, Enterococcus and Staphylococcus aureus. This capture was attributed to the aggregation of negatively charged nanoparticles on bacterial cell regional surfaces that bear positive charges. Three types of non-porous silica-encapsulated anion-exchange magnetic microparticles (SiMAG-Q, SiMAG-PEI, SiMAG-DEAE) were capable of concentrating a variety of bacteria, and were compared with silica-free, smaller fluidMAG particles. Salmonella, Escherichia coli, Enterococcus and other bacteria spiked in aqueous solutions, tap water and reservoir water were separated and concentrated by membrane filtration and magnetic separation based on these ion-exchange magnetic materials, and then characterized by whole cell MALDI-MS. By comparing with the mass spectra of the isolates and pure cells, bacteria in fresh water can be rapidly detected at 1 x 10(3) colony-forming units (cfu)/mL.

摘要

饮用水中微生物污染物的鉴定对基质辅助激光解吸电离质谱(MALDI-MS)来说是一个挑战,因为淡水中微生物的含量很低。为了避免为后续 MALDI-MS 分析获得足够微生物细胞的耗时培养步骤,膜过滤和基于纳米粒子或微粒的磁性分离的组合是一种快速有效的方法。在这项工作中,通过 MALDI-MS 分析和透射电子显微镜研究了细菌与流体 MAG-PAA(一种阳离子交换超顺磁纳米材料)的相互作用。FluidMAG-PAA 选择性地捕获了沙门氏菌、芽孢杆菌、肠球菌和金黄色葡萄球菌的细胞。这种捕获归因于带正电荷的细菌细胞区域表面上的带负电荷的纳米粒子的聚集。三种类型的无孔硅包裹的阴离子交换磁性微球(SiMAG-Q、SiMAG-PEI、SiMAG-DEAE)能够浓缩多种细菌,并与无硅、较小的流体 MAG 颗粒进行了比较。通过膜过滤和基于这些离子交换磁性材料的磁性分离,从水溶液、自来水和水库水中分离和浓缩了经过沙门氏菌、大肠杆菌、肠球菌等细菌污染的水,并通过全细胞 MALDI-MS 进行了表征。通过与分离株和纯细胞的质谱进行比较,可以在 1 x 10(3)菌落形成单位(cfu)/mL 的水平快速检测新鲜水中的细菌。

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