Division of Computational and Systems Medicine, Department of Surgery and Cancer, Imperial College London, South Kensington Campus, London, UK.
The Hamlyn Centre, Department of Computing, Faculty of Engineering, Imperial College London, South Kensington Campus, London, UK.
Sci Rep. 2019 Feb 28;9(1):3006. doi: 10.1038/s41598-019-39815-w.
The accurate and timely identification of the causative organism of infection is important in ensuring the optimum treatment regimen is prescribed for a patient. Rapid evaporative ionisation mass spectrometry (REIMS), using electrical diathermy for the thermal disruption of a sample, has been shown to provide fast and accurate identification of microorganisms directly from culture. However, this method requires contact to be made between the REIMS probe and microbial biomass; resulting in the necessity to clean or replace the probes between analyses. Here, optimisation and utilisation of ambient laser desorption ionisation (ALDI) for improved speciation accuracy and analytical throughput is shown. Optimisation was completed on 15 isolates of Escherichia coli, showing 5 W in pulsatile mode produced the highest signal-to-noise ratio. These parameters were used in the analysis of 150 clinical isolates from ten microbial species, resulting in a speciation accuracy of 99.4% - higher than all previously reported REIMS modalities. Comparison of spectral data showed high levels of similarity between previously published electrical diathermy REIMS data. ALDI does not require contact to be made with the sample during analysis, meaning analytical throughput can be substantially improved, and further, increases the range of sample types which can be analysed in potential direct-from-sample pathogen detection.
准确、及时地鉴定感染的病原体对于确保为患者开出最佳治疗方案非常重要。快速蒸发离解质谱(REIMS)利用电热来对样本进行热破坏,已被证明可直接从培养物中快速准确地鉴定微生物。然而,这种方法要求 REIMS 探头与微生物生物量接触;这就需要在分析之间清洁或更换探头。本文展示了优化和利用环境激光解吸电离(ALDI)以提高分类准确性和分析通量的方法。对 15 株大肠杆菌的优化表明,在脉冲模式下使用 5W 可产生最高的信噪比。这些参数用于分析来自十种微生物的 150 个临床分离株,分类准确性达到 99.4% - 高于所有以前报道的 REIMS 模式。光谱数据的比较表明,以前发表的电热 REIMS 数据之间具有高度相似性。ALDI 在分析过程中不需要与样本接触,这意味着分析通量可以大大提高,并且可以进一步增加可以在潜在的直接从样本中进行病原体检测中分析的样本类型范围。