Nanoxis AB, Lennart Torstenssonsgatan 5, SE-40016, Gothenburg, Sweden.
J Proteome Res. 2012 May 4;11(5):2710-20. doi: 10.1021/pr2010633. Epub 2012 Apr 11.
Because of the alarming expansion in the diversity and occurrence of bacteria displaying virulence and resistance to antimicrobial agents, it is increasingly important to be able to detect these microorganisms and to differentiate and identify closely related species, as well as different strains of a given species. In this study, a mass spectrometry proteomics approach is applied, exploiting lipid-based protein immobilization (LPI), wherein intact bacterial cells are bound, via membrane-gold interactions, within a FlowCell. The bound cells are subjected to enzymatic digestion for the generation of peptides, which are subsequently identified, using LC-MS. Following database matching, strain-specific peptides are used for subspecies-level discrimination. The method is shown to enable a reliable typing and identification of closely related strains of the same bacterial species, herein illustrated for Helicobacter pylori .
由于具有毒力和抗抗菌药物抗性的细菌的多样性和出现呈惊人的增长，因此能够检测这些微生物并区分和鉴定密切相关的物种以及给定物种的不同菌株变得越来越重要。在这项研究中，应用了一种质谱蛋白质组学方法，利用基于脂质的蛋白质固定化（LPI），其中完整的细菌细胞通过膜金相互作用结合在流池内。结合的细胞进行酶消化以生成肽，然后使用 LC-MS 进行鉴定。通过数据库匹配，使用亚种水平的区分来鉴定种内特定的肽。该方法被证明能够可靠地区分和鉴定相同细菌物种的密切相关菌株，本文以幽门螺杆菌为例进行了说明。
