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采用鸟枪法蛋白质组学和 LC-纳升电喷雾电离离子阱质谱联用技术诊断鲍曼不动杆菌的β-内酰胺类耐药性。

Diagnosis of β-lactam resistance in Acinetobacter baumannii using shotgun proteomics and LC-nano-electrospray ionization ion trap mass spectrometry.

机构信息

Department of Molecular Biology and Human Genetics, Tzu Chi University, Hualien 970, Taiwan.

出版信息

Anal Chem. 2013 Mar 5;85(5):2802-8. doi: 10.1021/ac303326a. Epub 2013 Feb 19.

Abstract

Acinetobacter baumannii is an important nosocomial pathogen that often affects critically ill patients in intensive care units. β-Lactam antibiotics are the most commonly prescribed drugs for infectious diseases caused by A. baumannii. Our aim is to develop an accurate and rapid shotgun proteomics method for the identification of β-lactam-resistant A. baumannii pathogens. In the present study, we used automated data-dependent scanning on a nano-LC/ion trap mass spectrometer to characterize proteotypic peptides of A. baumannii. Then, we used SEQUEST software to search specific databases, the β-lactam-resistance protein database of A. baumannii (BRPDAB). We successfully found a number of associated antibiotic-resistant proteins, including AmpC, β-lactamase, and carO, in clinical resistant strains of A. baumannii and differentiated them from wild-type A. baumannii strains. We used the results of the search to identify A. baumannii pathogens and found a β-lactam-resistant clinical strain of A. baumannii using Uniprot annotations, Gene Ontology (GO), and BLAST bioinformatics tools. This proteomic study will provide a platform for the rapid diagnosis of wild-type and resistant strains of A. baumannii, which would be useful for the medical treatment of these strains.

摘要

鲍曼不动杆菌是一种重要的医院获得性病原体,常影响重症监护病房的危重病患者。β-内酰胺类抗生素是治疗由鲍曼不动杆菌引起的感染性疾病最常用的药物。我们的目的是开发一种准确、快速的鸟枪法蛋白质组学方法,用于鉴定耐β-内酰胺的鲍曼不动杆菌病原体。在本研究中,我们使用自动数据依赖扫描技术在纳升 LC/离子阱质谱仪上对鲍曼不动杆菌的特征肽进行了分析。然后,我们使用 SEQUEST 软件搜索特定的数据库,即鲍曼不动杆菌的β-内酰胺耐药蛋白数据库(BRPDAB)。我们成功地在临床耐药株中找到了许多与抗生素耐药相关的蛋白,包括 AmpC、β-内酰胺酶和 carO,并将它们与野生型鲍曼不动杆菌株区分开来。我们使用搜索结果来鉴定鲍曼不动杆菌病原体,并使用 Uniprot 注释、基因本体论(GO)和 BLAST 生物信息学工具发现了一株耐β-内酰胺的临床鲍曼不动杆菌菌株。这项蛋白质组学研究将为快速诊断野生型和耐药型鲍曼不动杆菌提供一个平台,这对于这些菌株的治疗将非常有用。

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