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首页> 外文期刊>Journal of Clinical Microbiology >Detection of Colistin Resistance in Escherichia coli by Use of the MALDI Biotyper Sirius Mass Spectrometry System
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Detection of Colistin Resistance in Escherichia coli by Use of the MALDI Biotyper Sirius Mass Spectrometry System

机译：使用MALDI Biotyper Sirius质谱系统检测大肠杆菌中的共利斯汀抗性

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Polymyxin antibiotics are a last-line treatment for multidrug-resistant Gram-negative bacteria. However, the emergence of colistin resistance, including the spread of mobile mcr genes, necessitates the development of improved diagnostics for the detection of colistin-resistant organisms in hospital settings. The recently developed MALDIxin test enables detection of colistin resistance by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) in less than 15 min but is not optimized for the mass spectrometers commonly found in clinical microbiology laboratories. In this study, we adapted the MALDIxin test for the MALDI Biotyper Sirius MALDI-TOF MS system (Bruker Daltonics). We optimized the sample preparation protocol by using a set of 6 mobile colistin resistance (MCR) protein-expressing Escherichia coli clones and validated the assay with a collection of 40 E. coli clinical isolates, including 19 confirmed MCR protein producers, 12 colistin-resistant isolates that tested negative for commonly encountered mcr genes (i.e., likely chromosomally resistant isolates), and 9 polymyxin-susceptible isolates. We calculated polymyxin resistance ratio (PRR) values from the acquired spectra; PRR values of 0, indicating polymyxin susceptibility, were obtained for all colistin-susceptible E. coli isolates, whereas positive PRR values, indicating resistance to polymyxins, were obtained for all resistant strains, independent of the genetic basis of resistance. Thus, we report a preliminary feasibility study showing that an optimized version of the MALDIxin test adapted for the routine MALDI Biotyper Sirius system provides an unbiased, fast, reliable, cost-effective, and high-throughput way of detecting colistin resistance in clinical E. coli isolates.
机译：多粘菌素抗生素是耐多药革兰氏阴性菌的最后治疗方法。然而，大肠菌素抗性的出现，包括移动mcr基因的传播，使得有必要开发改进的诊断方法以检测医院环境中的大肠菌素抗性生物。最近开发的MALDIxin测试能够在不到15分钟的时间内通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）检测大肠菌素抗性，但并未针对临床微生物实验室中常见的质谱仪进行优化。在这项研究中，我们将MALDIxin测试用于MALDI Biotyper Sirius MALDI-TOF MS系统（布鲁克·道尔顿公司）。我们通过使用一组6个表达大肠杆菌表达的大肠埃希氏菌克隆优化了样品制备方案，并用40种大肠杆菌临床分离株进行了验证，包括19种已确证的MCR蛋白生产者，12种对大肠菌素具有耐药性对常见的mcr基因测试呈阴性的分离株（即可能具有抗染色体性的分离株）和9种对多粘菌素敏感的分离株。我们从获得的光谱中计算了多粘菌素抗性比（PRR）值;对于所有大肠杆菌敏感的大肠杆菌分离物，获得的PRR值为0，表明多粘菌素敏感性，而对于所有抗性菌株，获得的PRR值均为正，表明对多粘菌素具有抗性，而与抗药性的遗传基础无关。因此，我们报告了一项初步的可行性研究，结果表明，适用于常规MALDI Biotyper Sirius系统的MALDIxin测试的优化版本提供了一种无偏见，快速，可靠，具有成本效益的高通量方式来检测临床大肠杆菌中的大肠菌素抗性。大肠杆菌分离株。

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