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在赫尔辛基和新地区医院,用重复序列PCR对鲍曼不动杆菌克隆进行快速分子鉴定,并通过新型多重PCR评估碳青霉烯酶基因。

Rapid molecular characterization of Acinetobacter baumannii clones with rep-PCR and evaluation of carbapenemase genes by new multiplex PCR in Hospital District of Helsinki and Uusimaa.

作者信息

Pasanen Tanja, Koskela Suvi, Mero Sointu, Tarkka Eveliina, Tissari Päivi, Vaara Martti, Kirveskari Juha

机构信息

Division of Clinical Microbiology, Helsinki University Hospital, Helsinki, Finland.

出版信息

PLoS One. 2014 Jan 21;9(1):e85854. doi: 10.1371/journal.pone.0085854. eCollection 2014.

Abstract

Multidrug-resistant Acinetobacter baumannii (MDRAB) is an increasing problem worldwide. Prevalence of carbapenem resistance in Acinetobacter spp. due to acquired carbapenemase genes is not known in Finland. The purpose of this study was to examine prevalence and clonal spread of multiresistant A. baumannii group species, and their carbapenemase genes. A total of 55 Acinetobacter isolates were evaluated with repetitive PCR (DiversiLab) to analyse clonality of isolates, in conjunction with antimicrobial susceptibility profile for ampicillin/sulbactam, colistin, imipenem, meropenem, rifampicin and tigecycline. In addition, a new real-time PCR assay, detecting most clinically important carbapenemase genes just in two multiplex reactions, was developed. The assay detects genes for KPC, VIM, IMP, GES-1/-10, OXA-48, NDM, GIM-1, SPM-1, IMI/NMC-A, SME, CMY-10, SFC-1, SIM-1, OXA-23-like, OXA-24/40-like, OXA-58 and ISAbaI-OXA-51-like junction, and allows confident detection of isolates harbouring acquired carbapenemase genes. There was a time-dependent, clonal spread of multiresistant A. baumannii strongly correlating with carbapenamase gene profile, at least in this geographically restricted study material. The new carbapenemase screening assay was able to detect all the genes correctly suggesting it might be suitable for epidemiologic screening purposes in clinical laboratories.

摘要

多重耐药鲍曼不动杆菌(MDRAB)在全球范围内是一个日益严重的问题。在芬兰,不动杆菌属中由于获得性碳青霉烯酶基因导致的碳青霉烯耐药率尚不清楚。本研究的目的是检测多重耐药鲍曼不动杆菌群体物种的流行情况、克隆传播及其碳青霉烯酶基因。总共55株不动杆菌分离株通过重复PCR(DiversiLab)进行评估,以分析分离株的克隆性,并结合氨苄西林/舒巴坦、黏菌素、亚胺培南、美罗培南、利福平及替加环素的药敏谱。此外,还开发了一种新的实时PCR检测方法,仅通过两个多重反应就能检测出大多数临床上重要的碳青霉烯酶基因。该检测方法可检测KPC、VIM、IMP、GES-1/-10、OXA-48、NDM、GIM-1、SPM-1、IMI/NMC-A、SME、CMY-10、SFC-1、SIM-1、OXA-23样、OXA-24/40样、OXA-58及ISAbaI-OXA-51样连接区的基因,并能可靠地检测出携带获得性碳青霉烯酶基因的分离株。至少在这个地理范围有限的研究材料中,多重耐药鲍曼不动杆菌存在时间依赖性的克隆传播,且与碳青霉烯酶基因谱密切相关。新的碳青霉烯酶筛查检测方法能够正确检测出所有基因,表明它可能适用于临床实验室的流行病学筛查。

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