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使用DiversiLab基于重复序列的半自动聚合酶链反应对意大利不同医院分离的鲍曼不动杆菌进行流行病学分析。

Use of the DiversiLab semiautomated repetitive-sequence-based polymerase chain reaction for epidemiologic analysis on Acinetobacter baumannii isolates in different Italian hospitals.

作者信息

Carretto Edoardo, Barbarini Daniela, Farina Claudio, Grosini Alessia, Nicoletti Pierluigi, Manso Esther

机构信息

Laboratori Sperimentali di Ricerca, Area Infettivologica, Fondazione IRCCS Policlinico San Matteo, Pavia 27100, Italy.

出版信息

Diagn Microbiol Infect Dis. 2008 Jan;60(1):1-7. doi: 10.1016/j.diagmicrobio.2007.07.002. Epub 2007 Sep 21.

Abstract

Acinetobacter baumannii is typically a nosocomial pathogen. Epidemiologic tools that can rapidly trace the spread of hospital-associated infections due to this microorganism are essential. Currently, amplified fragment length polymorphism and pulsed-field gel electrophoresis using ApaI, a macrorestriction enzyme, are the molecular techniques most widely used to type this microorganism. Unfortunately, they are technically demanding, requiring also well-trained personnel, and are time consuming. The aims of this study are 1) to evaluate the usefulness of the semiautomated repetitive-sequence-based polymerase chain reaction (rep-PCR) for typing A. baumannii, comparing this method with another semiautomated technique, such as ribotyping, and 2) to acquire information about the incidence, the clinical significance, and the susceptibility patterns of this microorganism in 13 different Italian hospitals in a 4-week period (total study population, >14000 beds). Twenty-eight A. baumannii were isolated in 7 different hospitals; 21 strains were analyzed with molecular methods. Automated ribotyping distinguished 6 different clusters of isolates, whereas rep-PCR appeared to be more discriminating, allowing us to distinguish 8 different clusters. Our study confirms the good discriminatory power of the semiautomated rep-PCR. Although expensive, this method is simple, fast, and reproducible, and in our opinion, it could be used in a hierarchic approach as a 1st-line typing tool if results of analysis are required in a short period or if a large number of isolates have to be analyzed.

摘要

鲍曼不动杆菌通常是一种医院病原体。能够快速追踪由这种微生物引起的医院感染传播的流行病学工具至关重要。目前,扩增片段长度多态性和使用限制性内切酶ApaI的脉冲场凝胶电泳是最广泛用于对这种微生物进行分型的分子技术。不幸的是,它们技术要求高,并需要训练有素的人员,而且耗时。本研究的目的是:1)评估基于重复序列的半自动化聚合酶链反应(rep-PCR)对鲍曼不动杆菌进行分型的实用性,并将该方法与另一种半自动化技术(如核糖体分型)进行比较;2)获取有关这种微生物在4周内于意大利13家不同医院中的发病率、临床意义和药敏模式的信息(总研究人群超过14000张床位)。在7家不同医院分离出28株鲍曼不动杆菌;对其中21株菌株进行了分子方法分析。自动化核糖体分型区分出6个不同的分离株簇,而rep-PCR似乎更具鉴别力,可以区分出8个不同的簇。我们的研究证实了半自动化rep-PCR具有良好的鉴别力。尽管该方法成本高昂,但简单、快速且可重复,我们认为,如果需要在短时间内得到分析结果,或者需要分析大量分离株,它可以作为一线分型工具以分层方法使用。

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