Computational and Systems Medicine, Imperial College London , South Kensington Campus, London, SW7 2AZ, United Kingdom.
Imperial College Parturition Research Group, Institute of Reproductive and Developmental Biology, Imperial College London , London, W12 0NN, United Kingdom.
Anal Chem. 2017 Feb 7;89(3):1540-1550. doi: 10.1021/acs.analchem.6b03405. Epub 2017 Jan 11.
Medical swabs are routinely used worldwide to sample human mucosa for microbiological screening with culture methods. These are usually time-consuming and have a narrow focus on screening for particular microorganism species. As an alternative, direct mass spectrometric profiling of the mucosal metabolome provides a broader window into the mucosal ecosystem. We present for the first time a minimal effort/minimal-disruption technique for augmenting the information obtained from clinical swab analysis with mucosal metabolome profiling using desorption electrospray ionization mass spectrometry (DESI-MS) analysis. Ionization of mucosal biomass occurs directly from a standard rayon swab mounted on a rotating device and analyzed by DESI MS using an optimized protocol considering swab-inlet geometry, tip-sample angles and distances, rotation speeds, and reproducibility. Multivariate modeling of mass spectral fingerprints obtained in this way readily discriminate between different mucosal surfaces and display the ability to characterize biochemical alterations induced by pregnancy and bacterial vaginosis (BV). The method was also applied directly to bacterial biomass to confirm the ability to detect intact bacterial species from a swab. These results highlight the potential of direct swab analysis by DESI-MS for a wide range of clinical applications including rapid mucosal diagnostics for microbiology, immune responses, and biochemistry.
医用拭子在全球范围内被常规用于通过培养方法对人类黏膜进行微生物筛选取样。这些方法通常耗时较长,并且只能针对特定微生物物种进行筛选。作为替代方法,直接对黏膜代谢组进行质谱分析可以更全面地了解黏膜生态系统。我们首次提出了一种微创/微扰技术,通过解吸电喷雾电离质谱(DESI-MS)分析,在使用临床拭子分析获得的信息基础上,进一步进行黏膜代谢组分析。将标准的人造纤维拭子安装在旋转装置上,直接对黏膜生物量进行离子化,通过 DESI-MS 分析,并根据拭子入口几何形状、尖端-样品角度和距离、旋转速度以及重现性来优化分析方案。以这种方式获得的质谱指纹图谱的多元建模可以轻松区分不同的黏膜表面,并显示出能够表征妊娠和细菌性阴道病(BV)引起的生化变化的能力。该方法还直接应用于细菌生物量,以确认从拭子中检测完整细菌物种的能力。这些结果突出了直接通过 DESI-MS 对拭子进行分析的广泛临床应用潜力,包括用于微生物学、免疫反应和生物化学的快速黏膜诊断。