Analysis of Bacteriophages with Insulator-Based Dielectrophoresis
Analysis of Bacteriophages with Insulator-Based Dielectrophoresis
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DOI：
10.3390/mi10070450
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发表时间：
2019-07-01
期刊：
影响因子：
3.4
通讯作者：
Lapizco-Encinas, Blanca H.
中科院分区：
文献类型：
--
作者：
De Pena, Adriana Coll;Redzuan, Nurul Humaira Mohd;Lapizco-Encinas, Blanca H.
细菌病毒或病原体在医学和农业领域作为抗生素的替代物以控制致病细菌的有害种群具有巨大的潜力。然而，目前用于噬菌体的分析和纯化方案往往是资源密集型的，并且具有许多限制，例如影响噬菌体活力。本研究探讨了采用基于绝缘体的介电电泳（iDEP）的电动技术用于病毒评估，分离和富集的潜力。特别地，探索参数“捕获值”（Tv）的应用作为每个噬菌体物种的标准化iDEP签名。本研究包括COMSOL Multiphysics的数学建模和广泛的实验。研究了三种相关但遗传和结构不同的噬菌体：肠道沙门氏菌噬菌体SPN 3US、铜绿假单胞菌噬菌体phi KZ和绿针假单胞菌噬菌体201 phi 2-1。这是第一个对具有大而复杂的病毒体的噬菌体进行的iDEP研究，结果表明它们的病毒体可以成功地用iDEP系统富集，并且仍然保持感染性。此外，我们的研究结果表明，表征的负介电电泳反应的噬菌体在Tv可用于预测个别病毒的行为在iDEP系统。这里报告的研究结果可以有助于建立协议，分析，纯化和/或富集已知和未知的细菌样品。
Bacterial viruses or phages have great potential in the medical and agricultural fields as alternatives to antibiotics to control nuisance populations of pathogenic bacteria. However, current analysis and purification protocols for phages tend to be resource intensive and have numbers of limitations, such as impacting phage viability. The present study explores the potential of employing the electrokinetic technique of insulator-based dielectrophoresis (iDEP) for virus assessment, separation and enrichment. In particular, the application of the parameter "trapping value" (Tv) is explored as a standardized iDEP signature for each phage species. The present study includes mathematical modeling with COMSOL Multiphysics and extensive experimentation. Three related, but genetically and structurally distinct, phages were studied: Salmonella enterica phage SPN3US, Pseudomonas aeruginosa phage phi KZ and P. chlororaphis phage 201 phi 2-1. This is the first iDEP study on bacteriophages with large and complex virions and the results illustrate their virions can be successfully enriched with iDEP systems and still retain infectivity. In addition, our results indicate that characterization of the negative dielectrophoretic response of a phage in terms of Tv could be used for predicting individual virus behavior in iDEP systems. The findings reported here can contribute to the establishment of protocols to analyze, purify and/or enrich samples of known and unknown phages.