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一种基于基质辅助激光解吸电离飞行时间质谱的全面方法来检测产质粒编码KPC的…… (原文不完整)

A Full MALDI-Based Approach to Detect Plasmid-Encoded KPC-Producing .

作者信息

Cordovana Miriam, Kostrzewa Markus, Glandorf Jörg, Bienia Michael, Ambretti Simone, Pranada Arthur B

机构信息

Laboratory of Bacteriology, Operative Unit of Microbiology, University Hospital of Bologna Policlinico Sant'Orsola-Malpighi, Bologna, Italy.

Bruker Daltonik GmbH, Bremen, Germany.

出版信息

Front Microbiol. 2018 Nov 23;9:2854. doi: 10.3389/fmicb.2018.02854. eCollection 2018.

Abstract

KPC-producing represents a severe public health concern worldwide. The rapid detection of these isolates is of fundamental importance for the adoption of proper antibiotic treatment and infection control measures, and new applications of MALDI-TOF MS technology fit this purpose. In this study, we present a full MALDI-based approach to detect plasmid-encoded KPC-producing strains, accomplished by the automated detection of a KPC-specific peak (at 11,109 m/z) by a specific algorithm integrated into the MALDI Biotyper system (Bruker Daltonik), and the confirmation of carbapenemase activity by STAR-Carba imipenem hydrolysis assay. A total of 6209 isolates from Italy and Germany were investigated for the presence of the KPC-related peak, and a subset of them ( = 243) underwent confirmation of carbapenemase activity by STAR-Carba assay. The novel approach was further applied directly to positive blood culture bottles ( = 204), using the bacterial pellet obtained with Sepsityper kit (Bruker Daltonik). The novel approach enabled a reliable and very fast detection of KPC-producing strains, from colonies as well as directly from positive blood cultures. The automated peak detection enabled the instant detection of KPC-producing during the routine identification process, with excellent specificity (100%) and a good sensitivity (85.1%). The sensitivity is likely mainly related to the prevalence of the specific plasmid harboring clones among all the KPC-producing circulating strains. STAR-Carba carbapenemase confirmation showed 100% sensitivity and specificity, both from colonies and from positive blood cultures.

摘要

产KPC酶的菌株是全球严重的公共卫生问题。快速检测这些分离株对于采取适当的抗生素治疗和感染控制措施至关重要,而基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)技术的新应用符合这一目的。在本研究中,我们提出了一种基于MALDI的完整方法来检测携带质粒的产KPC酶菌株,该方法通过整合到MALDI Biotyper系统(布鲁克道尔顿公司)中的特定算法自动检测KPC特异性峰(质荷比为11,109),并通过STAR-Carba亚胺培南水解试验确认碳青霉烯酶活性。对来自意大利和德国的6209株分离株进行了KPC相关峰的检测,其中一部分(n = 243)通过STAR-Carba试验确认了碳青霉烯酶活性。该新方法还直接应用于阳性血培养瓶(n = 204),使用Sepsityper试剂盒(布鲁克道尔顿公司)获得的细菌沉淀。这种新方法能够可靠且快速地检测出产KPC酶的菌株,无论是从菌落还是直接从阳性血培养物中。自动峰检测能够在常规鉴定过程中即时检测出产KPC酶的菌株,具有出色的特异性(100%)和良好的敏感性(85.1%)。敏感性可能主要与所有产KPC酶的流行菌株中携带特定质粒的克隆的流行率有关。STAR-Carba碳青霉烯酶确认试验显示,无论是从菌落还是从阳性血培养物中,敏感性和特异性均为100%。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1c04/6277887/adee645c61e1/fmicb-09-02854-g001.jpg

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