Moscow Institute of Physics and Technology, Dolgoprudny, Russia.
Moscow Institute of Physics and Technology, Dolgoprudny, Russia; Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of Russian Academy of Sciences, Moscow, Russia.
Biosens Bioelectron. 2022 Oct 1;213:114444. doi: 10.1016/j.bios.2022.114444. Epub 2022 Jun 1.
With the rise of drug resistance, bacteriophages and bacteriophage-derived proteins may become an efficient successor to traditional antibiotics. While the enormous natural diversity of the phages allows matching virtually any bacteria, identification of the potentially life-saving phage is currently a tedious and time-consuming challenge that often cannot be performed within a reasonable time. Here we show a rapid 1-min bacteriophage screening assay based on specially constructed phage-mimicking nanoagents and surface plasmon resonance effect. Within the assay, a panel of phage-mimicking gold nanoparticles, possessing the specificity and enzymatic activity of a particular phage, is mixed with a suspension of the bacteria of interest. The spectral behaviour of the assay mix allows measurement of two critical parameters of the nanoagents and the corresponding bacteriophages: 1) direct assessment of their specificity due to convergence of the particles on the cell walls, and more importantly, 2) real-time evaluation of their enzymatic activity for the destruction of the cell capsule via detection of nanoagent detachment from the surface of bacteria. The proposed assay overcomes the current time limitations of the phage-bacteria matching procedures and thereby can facilitate faster development and adoption of phage-based therapies as a much-needed alternative to traditional antibiotics.
随着耐药性的出现，噬菌体和噬菌体衍生蛋白可能成为传统抗生素的有效替代品。虽然噬菌体具有巨大的自然多样性，可以匹配几乎任何细菌，但目前识别潜在救命噬菌体是一项繁琐且耗时的挑战，往往无法在合理的时间内完成。在这里，我们展示了一种基于特殊构建的噬菌体模拟纳米制剂和表面等离子体共振效应的快速 1 分钟噬菌体筛选测定法。在该测定法中，一组具有特定噬菌体特异性和酶活性的噬菌体模拟金纳米粒子与感兴趣的细菌悬浮液混合。测定法混合物的光谱行为允许测量纳米制剂和相应噬菌体的两个关键参数：1）由于粒子在细胞壁上的收敛，直接评估它们的特异性，更重要的是，2）通过检测纳米制剂从细菌表面的脱离，实时评估它们破坏细胞胶囊的酶活性。所提出的测定法克服了当前噬菌体-细菌匹配程序的时间限制，从而可以加快噬菌体治疗的开发和采用，作为对传统抗生素的急需替代。
