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Cationized magnetoferritin enables rapid labeling and concentration of Gram-positive and Gram-negative bacteria in magnetic cell separation columns

机译:阳离子化的铁铁蛋白可在磁性细胞分离柱中快速标记和浓缩革兰氏阳性和阴性细菌

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摘要

In order to identify pathogens rapidly and reliably, it is often required to capture and concentrate them from large sample volumes into smaller ones. Magnetic labeling and capture of bacteria using a magnetic field holds great promise towards achieving this goal, but current protocols suffer from poor capture efficiency. Here, we present a rapid and highly efficient approach to magnetic labeling and capture of both Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria using cationized magnetoferritin (cat- MF). Magnetic labeling was achieved within a one minute incubation period with cat-MF, and 99.97% of the labeled bacteria could be immobilized in commercially available magnetic cell separation (MACS) columns. Longer incubation times led to more efficient capture, with S. aureus being immobilized to a greater extent than E. coli. Finally, low numbers of magnetically labeled E. coli ( 100 colony forming units per mL) were immobilized with 100% efficiency and concentrated 7-fold within 15 minutes. Therefore, our study provides a novel protocol for rapid and highly efficient magnetic labeling, capture and concentration of both Gram-positive and Gram-negative bacteria.
机译:为了快速,可靠地识别病原体,通常需要将其从大样本量中捕获并浓缩成较小的样本。使用磁场进行磁性标记和细菌捕获对于实现该目标具有广阔的前景,但是目前的方案存在捕获效率差的问题。在这里,我们介绍了一种快速高效的方法,用于使用阳离子化的磁铁蛋白(cat-MF)进行革兰氏阴性(Escherichia coli)和革兰氏阳性(Staphylococcus aureus)细菌的磁性标记和捕获。在与cat-MF孵育一分钟内完成磁性标记,并且99.97%的标记细菌可以固定在市售的磁性细胞分离(MACS)色谱柱中。更长的孵育时间导致更有效的捕获,与大肠杆菌相比,金黄色葡萄球菌的固定程度更高。最后,以100%的效率固定了少量的磁性标记大肠杆菌(每毫升<100个菌落形成单位),并在15分钟内浓缩了7倍。因此,我们的研究为革兰氏阳性菌和革兰氏阴性菌的快速,高效的磁性标记,捕获和浓缩提供了一种新颖的方案。

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