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通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)快速检测源自血培养的产碳青霉烯酶肺炎克雷伯菌菌株。

Rapid detection of carbapenemase-producing Klebsiella pneumoniae strains derived from blood cultures by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS).

作者信息

Sakarikou Christina, Ciotti Marco, Dolfa Camilla, Angeletti Silvia, Favalli Cartesio

机构信息

Department of Experimental Medicine and Surgery, "Tor Vergata" University of Rome, Via Montpellier 1, 00133, Rome, Italy.

Laboratory of Clinical Microbiology and Virology, Polyclinic "Tor Vergata" Foundation, V.le Oxford 81, 00133, Rome, Italy.

出版信息

BMC Microbiol. 2017 Mar 8;17(1):54. doi: 10.1186/s12866-017-0952-3.

Abstract

BACKGROUND

Carbapenemase-producing Enterobacteriaceae (CPE), particularly carbapenemase-producing Klebsiella pneumoniae isolates, are important causative agents of nosocomial infections associated with significant mortality rates mostly in critical wards. The rapid detection and typing of these strains is critical either for surveillance purposes and to prevent outbreaks and optimize antibiotic therapy. In this study, the MALDI-TOF MS method was used to detect rapidly these isolates from blood cultures (BCs) and to obtain proteomic profiles enable to discriminate between carbapenemase-producing and non-carbapenemase-producing strains.

RESULTS

Fifty-five K. pneumoniae strains were tested. Identification and carbapenemase-production detection assay using Ertapenem were performed both from bacterial pellets extracted directly from BCs flasks and from subcultures of these strains. For all isolates, a complete antimicrobial susceptibility testing and a genotypic characterization were performed. We found 100% agreement between the carbapenemase-producing profile generated by MALDI TOF MS and that obtained using conventional methods. The assay detected and discriminated different carbapenemase-producing K. pneumoniae isolates within 30 min to 3 h after incubation with Ertapenem.

CONCLUSIONS

MALDI-TOF MS is a promising, rapid and economical method for the detection of carbapenemase-producing K. pneumoniae strains that could be successfully introduced into the routine diagnostic workflow of clinical microbiology laboratories.

摘要

背景

产碳青霉烯酶肠杆菌科细菌(CPE),尤其是产碳青霉烯酶肺炎克雷伯菌分离株,是医院感染的重要病原体,主要在重症监护病房导致较高的死亡率。快速检测和分型这些菌株对于监测、预防暴发以及优化抗生素治疗都至关重要。在本研究中,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)方法被用于从血培养物(BCs)中快速检测这些分离株,并获得能够区分产碳青霉烯酶菌株和非产碳青霉烯酶菌株的蛋白质组图谱。

结果

对55株肺炎克雷伯菌菌株进行了检测。使用厄他培南进行的鉴定和碳青霉烯酶产生检测试验,既从直接从血培养瓶中提取的细菌沉淀进行,也从这些菌株的传代培养物进行。对所有分离株进行了完整的抗菌药物敏感性测试和基因分型鉴定。我们发现MALDI TOF MS生成的产碳青霉烯酶图谱与使用传统方法获得的图谱完全一致。该试验在与厄他培南孵育后3小时内即可检测并区分不同的产碳青霉烯酶肺炎克雷伯菌分离株。

结论

MALDI-TOF MS是一种有前景、快速且经济的检测产碳青霉烯酶肺炎克雷伯菌菌株的方法,可成功引入临床微生物学实验室的常规诊断工作流程。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3a69/5343375/6474ddd62f53/12866_2017_952_Fig1_HTML.jpg

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