School of Chemistry and Manchester Institute of Biotechnology, University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.
Analyst. 2013 Mar 7;138(5):1363-9. doi: 10.1039/c3an36517d.
Fourier transform infrared (FT-IR) spectroscopy is an established rapid whole-organism fingerprinting method that generates metabolic fingerprints from bacteria that reflect the phenotype of the microorganism under investigation. However, whilst FT-IR spectroscopy is fast (typically 10 s to 1 min per sample), the approaches for microbial sample preparation can be time consuming as plate culture or shake flasks are used for growth of the organism. We report a new approach that allows micro-cultivation of bacteria from low volumes (typically 200 μL) to be coupled with FT-IR spectroscopy. This approach is fast and easy to perform and gives equivalent data to the lengthier and more expensive shake flask cultivations (sample volume = 20 mL). With this micro-culture approach we also demonstrate high reproducibility of the metabolic fingerprints. The approach allowed separation of different isolates of Escherichia coli involved in urinary tract infection, including members of the globally disseminated ST131 clone, with respect to both genotype and resistance or otherwise to the antibiotic Ciprofloxacin.
傅里叶变换红外(FT-IR)光谱是一种成熟的快速全生物体指纹识别方法,它从细菌中生成代谢指纹,反映出被研究微生物的表型。然而,尽管 FT-IR 光谱速度很快(通常每个样品为 10 秒至 1 分钟),但微生物样品制备的方法可能很耗时,因为需要使用平板培养或摇瓶来培养微生物。我们报告了一种新方法,可将从低体积(通常为 200 μL)中培养的细菌与 FT-IR 光谱相结合。这种方法快速且易于操作,并可获得与耗时更长且更昂贵的摇瓶培养(样品体积=20 mL)相当的数据。通过这种微培养方法,我们还证明了代谢指纹具有很高的重现性。该方法能够分离与尿路感染有关的不同大肠杆菌分离株,包括在全球传播的 ST131 克隆的成员,同时能够区分它们的基因型以及对环丙沙星抗生素的耐药性或敏感性。