School of Molecular Sciences, Arizona State University, Tempe, AZ 85287, USA..
Analyst. 2020 Apr 21;145(8):2904-2914. doi: 10.1039/c9an01449g. Epub 2020 Feb 19.
Differentiating bacteria strains using biophysical forces has been the focus of recent studies using dielectrophoresis (DEP). The refinement of these studies has created high-resolution separations such that very subtle properties of the cells are enough to induce significant differences in measurable biophysical properties. These high-resolution capabilities build upon the advantages of DEP which include small sample sizes and fast analysis times. Studies focusing on differentiating antimicrobial resistant and susceptible bacteria potentially have significant impact on human health and medical care. A prime example is Staphylococcus aureus, which commonly colonizes adults without ill effects. However, the pathogen is an important cause of infections, including surgical site infections. Treatment of S. aureus infections is generally possible with antimicrobials, but antimicrobial resistance has emerged. Of special importance is resistance to methicillin, an antimicrobial created in response to resistance to penicillin. Here, dielectrophoresis is used to study methicillin-resistant (MRSA) and -susceptible S. aureus (MSSA) strains, both with and without the addition of a fluorescent label. The capture onset potential of fluorescently-labeled MRSA (865 ± 71 V) and thus the ratio of electrokinetic to dielectrophoretic mobility, was found to be higher than that of fluorescently-labeled MSSA (685 ± 61 V). This may be attributable to the PBP2a enzyme present in the MRSA strain and not in the MSSA bacteria. Further, unlabeled MRSA was found to have a capture onset potential of 732 ± 44 V, while unlabeled MSSA was found to have a capture onset potential of 562 ± 59 V. This shows that the fluorescently-labeled bacteria require a higher applied potential, and thus ratio of mobilities, to capture than the unlabeled bacteria.
使用介电泳(DEP)区分细菌菌株一直是最近研究的重点。这些研究的精细化已经实现了高分辨率分离,使得细胞的非常细微特性足以在可测量的生物物理特性中引起显著差异。这些高分辨率能力建立在DEP 的优势之上,包括小样本量和快速分析时间。专注于区分抗微生物药物敏感和耐药细菌的研究有可能对人类健康和医疗保健产生重大影响。一个主要的例子是金黄色葡萄球菌,它通常在成年人中定植而没有不良影响。然而,该病原体是感染的重要原因,包括手术部位感染。金黄色葡萄球菌感染的治疗通常可以使用抗生素,但抗生素耐药性已经出现。特别重要的是对甲氧西林的耐药性,甲氧西林是一种针对青霉素耐药性而开发的抗生素。在这里,使用介电泳来研究耐甲氧西林金黄色葡萄球菌(MRSA)和甲氧西林敏感金黄色葡萄球菌(MSSA)菌株,包括添加和不添加荧光标记的情况。荧光标记的 MRSA(865 ± 71 V)的捕获起始势,以及因此电泳和介电泳迁移率的比值,被发现高于荧光标记的 MSSA(685 ± 61 V)。这可能归因于 MRSA 菌株中存在而 MSSA 细菌中不存在的 PBP2a 酶。此外,发现未标记的 MRSA 的捕获起始势为 732 ± 44 V,而未标记的 MSSA 的捕获起始势为 562 ± 59 V。这表明荧光标记的细菌比未标记的细菌需要更高的施加电势,因此需要更高的迁移率比来捕获。