Centre for Bacterial Resistance Biology, Department of Life Sciences, Imperial College London, London, SW7 2AZ, UK.
Department of Bacteriology-Hygiene, Bicêtre Hospital, Assistance Publique-Hôpitaux de Paris, Le Kremlin-Bicêtre, France.
Eur J Clin Microbiol Infect Dis. 2023 Jun;42(6):669-679. doi: 10.1007/s10096-023-04587-9. Epub 2023 Mar 27.
Colistin is a last resort drug for the treatment of multiple drug-resistant (MDR) Gram-negative bacterial infections. Rapid methods to detect resistance are highly desirable. Here, we evaluated the performance of a commercially available MALDI-TOF MS-based assay for colistin resistance testing in Escherichia coli at two different sites. Ninety clinical E. coli isolates were provided by France and tested in Germany and UK using a MALDI-TOF MS-based colistin resistance assay. Lipid A molecules of the bacterial cell membrane were extracted using the MBT Lipid Xtract Kit™ (RUO; Bruker Daltonics, Germany). Spectra acquisition and evaluation were performed by the MBT HT LipidART Module of MBT Compass HT (RUO; Bruker Daltonics) on a MALDI Biotyper® sirius system (Bruker Daltonics) in negative ion mode. Phenotypic colistin resistance was determined by broth microdilution (MICRONAUT MIC-Strip Colistin, Bruker Daltonics) and used as a reference. Comparing the results of the MALDI-TOF MS-based colistin resistance assay with the data of the phenotypic reference method for the UK, sensitivity and specificity for the detection of colistin resistance were 97.1% (33/34) and 96.4% (53/55), respectively. Germany showed 97.1% (33/34) sensitivity and 100% (55/55) specificity for the detection of colistin resistance by MALDI-TOF MS. Applying the MBT Lipid Xtract™ Kit in combination with MALDI-TOF MS and dedicated software showed excellent performances for E. coli. Analytical and clinical validation studies must be performed to demonstrate the performance of the method as a diagnostic tool.
黏菌素是治疗多重耐药(MDR)革兰氏阴性细菌感染的最后手段。快速检测耐药性的方法非常需要。在这里,我们评估了一种市售的 MALDI-TOF MS 基方法在两个不同地点用于检测大肠埃希菌中黏菌素耐药性的性能。从法国提供了 90 株临床大肠埃希菌分离株,并在德国和英国使用 MALDI-TOF MS 基黏菌素耐药性检测方法进行测试。使用 MBT Lipid Xtract Kit™(RUO;Bruker Daltonics,德国)提取细菌细胞膜的脂 A 分子。通过 MBT Compass HT 上的 MBT HT LipidART 模块(RUO;Bruker Daltonics)在 MALDI Biotyper® sirius 系统(Bruker Daltonics)上以负离子模式进行光谱采集和评估。通过肉汤微量稀释法(MICRONAUT MIC-Strip Colistin,Bruker Daltonics)测定表型黏菌素耐药性,并作为参考。将 MALDI-TOF MS 基黏菌素耐药性检测结果与英国的表型参考方法的数据进行比较,检测黏菌素耐药性的敏感性和特异性分别为 97.1%(33/34)和 96.4%(53/55)。德国通过 MALDI-TOF MS 检测黏菌素耐药性的敏感性为 97.1%(33/34),特异性为 100%(55/55)。应用 MBT Lipid Xtract Kit 结合 MALDI-TOF MS 和专用软件,对大肠埃希菌的表现出色。必须进行分析和临床验证研究,以证明该方法作为诊断工具的性能。