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一种市售 MALDI-TOF MS 检测大肠埃希菌多粘菌素耐药性方法的双位点研究。

Two-site study on performances of a commercially available MALDI-TOF MS-based assay for the detection of colistin resistance in Escherichia coli.

机构信息

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.

出版信息

Eur J Clin Microbiol Infect Dis. 2023 Jun;42(6):669-679. doi: 10.1007/s10096-023-04587-9. Epub 2023 Mar 27.

Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/542d/10172212/cda18aed07f5/10096_2023_4587_Fig1_HTML.jpg

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