Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich Schiller University Jena, Helmholtzweg 4, 07743, Jena, Germany.
InfectoGnostics Research Campus Jena, Philosophenweg 7, 07743, Jena, Germany.
Anal Bioanal Chem. 2019 Aug;411(21):5445-5454. doi: 10.1007/s00216-019-01918-8. Epub 2019 May 31.
Bacteremia with its high mortality is a frequent case in clinical health care. Further, bacteremia includes the considerable risk of progressing to a sepsis. Even in case of survival, sepsis still entails diminished quality of life for the survivors and high indirect cost for the society. The crucial factor in sepsis is time. Therefore, timely description of adequate antibiotics is vital to reduce mortality and improve quality of life after survival. Despite that, the current gold standard of clinical bacteria diagnostic is based on cultivation of bacteria, which requires an average of 13-h cultivation. Consequently, there is a necessity for culture free identification methods without sacrificing the range of bacteria strains which can be identified. Raman microspectroscopy in general requires only single bacteria cells and has proven to offer high identification accuracies. However, the prerequisite for Raman microspectroscopy is a suitable isolation strategy to obtain single unharmed bacteria cells free from matrix. Moreover, in blood, bacteria are outnumbered by billions of blood cells. In this study, we present an isolation strategy to recover single bacteria cells from blood and evaluate their suitability for Raman microspectroscopic identification. Graphical abstract.
菌血症具有较高的死亡率，是临床医疗中的常见病例。此外，菌血症存在着发展为败血症的重大风险。即使幸存下来，败血症仍会降低幸存者的生活质量，并给社会带来高昂的间接成本。败血症的关键因素是时间。因此，及时使用适当的抗生素是降低死亡率和提高幸存者生活质量的关键。尽管如此，目前临床细菌诊断的金标准仍然基于细菌培养，这需要平均 13 小时的培养时间。因此，需要有一种无需牺牲可识别细菌菌株范围的无培养鉴定方法。拉曼显微镜通常只需要单个细菌细胞，并且已被证明具有很高的鉴定准确性。然而，拉曼显微镜的前提是需要一种合适的分离策略，以获得单个未受损的、无基质的细菌细胞。此外，在血液中，细菌的数量是数十亿个血细胞的数亿倍。在这项研究中，我们提出了一种从血液中回收单个细菌细胞的分离策略，并评估了它们用于拉曼光谱鉴定的适用性。
