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Rapid detection of colistin resistance in Acinetobacter baumannii using MALDI-TOF-based lipidomics on intact bacteria

机译:使用基于MALDI-TOF的脂质组学对完整细菌快速检测鲍曼不动杆菌中的大肠菌素抗性

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With the dissemination of extremely drug resistant bacteria, colistin is now considered as the last-resort therapy for the treatment of infection caused by Gram-negative bacilli (including carbapenemase producers). Unfortunately, the increase use of colistin has resulted in the emergence of resistance as well. In A. baumannii, colistin resistance is mostly?caused by the addition of phosphoethanolamine to the lipid A through the action of a phosphoethanolamine transferase chromosomally-encoded by the pmrC gene, which is regulated by the two-component system PmrA/PmrB. In A. baumannii clinical isolate the main resistance mechanism to colistin involves mutations in pmrA, pmrB or pmrC genes leading to the overexpression of pmrC. Although, rapid detection of resistance is one of the key issues to improve the treatment of infected patient, detection of colistin resistance in A. baumannii still relies on MIC determination through microdilution, which is time-consuming (16–24?h). Here, we evaluated the performance of a recently described MALDI-TOF-based assay, the MALDIxin test, which allows the rapid detection of colistin resistance-related modifications to lipid A (i.e phosphoethanolamine addition). This test accurately detected all colistin-resistant A. baumannii isolates in less than 15?minutes, directly on intact bacteria with a very limited sample preparation prior MALDI-TOF analysis.
机译:随着极强耐药菌的传播,大肠菌素现在被认为是治疗革兰氏阴性杆菌(包括碳青霉烯酶生产者)引起的感染的最后手段。不幸的是,粘菌素的使用增加也导致了耐药性的出现。在鲍曼不动杆菌中,大肠粘菌素抗性主要是由于在由pmrC基因进行染色体编码的磷酸乙醇胺转移酶的作用下向脂质A中添加了磷酸乙醇胺而引起的,而磷酸乙醇胺转移酶是由两组分系统PmrA / PmrB调节的。在鲍曼不动杆菌临床分离物中,对粘菌素的主要抗性机制涉及导致pmrC过表达的pmrA,pmrB或pmrC基因突变。尽管快速检测耐药性是改善感染患者治疗的关键问题之一,但鲍曼不动杆菌中大肠菌素耐药性的检测仍然依赖于通过微稀释测定MIC,这很费时(16–24?h)。在这里,我们评估了最近描述的基于MALDI-TOF的检测方法MALDIxin检测的性能,该检测可快速检测大肠杆菌素对脂质A的抗性相关修饰(即添加磷酸乙醇胺)。该测试可在不到15分钟的时间内直接在完整细菌上准确检测出所有对大肠菌素耐药的鲍曼不动杆菌菌株,并在进行MALDI-TOF分析之前使用非常有限的样品制备方法。

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