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使用自动化重复基因外回文序列 - 聚合酶链反应DNA指纹图谱和基质辅助激光解吸/电离飞行时间质谱法对嗜水气单胞菌临床分离株进行分型

Typing of Ochrobactrum anthropi clinical isolates using automated repetitive extragenic palindromic-polymerase chain reaction DNA fingerprinting and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry.

作者信息

Quirino Angela, Pulcrano Giovanna, Rametti Linda, Puccio Rossana, Marascio Nadia, Catania Maria Rosaria, Matera Giovanni, Liberto Maria Carla, Focà Alfredo

机构信息

Institute of Microbiology, Department of Health Sciences, "Magna Graecia" University, Viale Europa, Catanzaro, Italy.

出版信息

BMC Microbiol. 2014 Mar 22;14:74. doi: 10.1186/1471-2180-14-74.

Abstract

BACKGROUND

Ochrobactrum anthropi (O. anthropi), is a non-fermenting gram-negative bacillus usually found in the environment. Nevertheless, during the past decade it has been identified as pathogenic to immunocompromised patients. In this study, we assessed the usefulness of the automated repetitive extragenic palindromic-polymerase chain reaction (rep-PCR-based DiversiLab™ system, bioMèrieux, France) and of matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF MS) for typing of twentythree O. anthropi clinical isolates that we found over a four-months period (from April 2011 to August 2011) in bacteriemic patients admitted in the same operative unit of our hospital. Pulsed-field gel electrophoresis (PFGE), commonly accepted as the gold standard technique for typing, was also used. Analysis was carried out using the Pearson correlation coefficient to determine the distance matrice and the unweighted pair group method with arithmetic mean (UPGMA) to generate dendogram.

RESULTS

Rep-PCR analysis identified four different patterns: three that clustered together with 97% or more pattern similarity, and one whose members showed < 95% pattern similarity. Interestingly, strains isolated later (from 11/06/2011 to 24/08/2011) displayed a pattern with 99% similarity. MALDI-TOF MS evaluation clustered the twentythree strains of O. anthropi into a single group containing four distinct subgroups, each comprising the majority of strains clustering below 5 distance levels, indicating a high similarity between the isolates.

CONCLUSIONS

Our results indicate that these isolates are clonally-related and the methods used afforded a valuable contribution to the epidemiology, prevention and control of the infections caused by this pathogen.

摘要

背景

嗜水气单胞菌(O. anthropi)是一种通常存在于环境中的非发酵革兰氏阴性杆菌。然而,在过去十年中,它已被确定为对免疫功能低下患者具有致病性。在本研究中,我们评估了自动化重复基因外回文序列 - 聚合酶链反应(基于rep-PCR的DiversiLab™系统,法国生物梅里埃公司)和基质辅助激光解吸/电离飞行时间(MALDI-TOF MS)对我们在四个月期间(2011年4月至2011年8月)在我院同一手术科室收治的菌血症患者中发现的23株嗜水气单胞菌临床分离株进行分型的实用性。脉冲场凝胶电泳(PFGE),通常被认为是分型的金标准技术,也被使用。使用Pearson相关系数进行分析以确定距离矩阵,并使用算术平均的非加权配对组方法(UPGMA)生成树状图。

结果

Rep-PCR分析确定了四种不同的模式:三种模式聚类在一起,模式相似性为97%或更高,一种模式的成员显示出<95%的模式相似性。有趣的是,后来分离的菌株(2011年6月11日至2011年8月24日)显示出99%相似性的模式。MALDI-TOF MS评估将23株嗜水气单胞菌菌株聚为一组,该组包含四个不同的亚组,每个亚组包含大多数在5个距离水平以下聚类的菌株,表明分离株之间具有高度相似性。

结论

我们的结果表明这些分离株具有克隆相关性,所使用的方法为该病原体引起的感染的流行病学、预防和控制做出了有价值的贡献。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8861/3977698/11ed005458fd/1471-2180-14-74-1.jpg

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