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利用随机扩增多态性DNA分析对嗜水气单胞菌菌株进行克隆鉴定。

Clonal identification of Aeromonas hydrophila strains using randomly amplified polymorphic DNA analysis.

作者信息

Talon D, Mulin B, Thouverez M

机构信息

Institut des Sciences et des Techniques de l'Environnement, Université de Franche-Comté, Besançon, France.

出版信息

Eur J Epidemiol. 1998 Apr;14(3):305-10. doi: 10.1023/a:1007441019821.

Abstract

The suitability of arbitrary primer polymerase chain reaction (RAPD) as a typing technique was evaluated by comparing it with pulsed-field gel electrophoresis (PFGE) to characterize Aeromonas hydrophila strains isolated from a cluster of hospital-acquired infections. Five isolates from patients and 10 isolates from the water supply were compared to 10 epidemiologically unrelated strains isolated from patients and rivers. Two methods were used to prepare DNA and two primers (AP3 and AP5) were selected. The discriminatory power was better with the extractive DNA preparation than the boiling method. The discrimination of closely related from less related strains by PCR using AP3 was consistent with that by PFGE: water supply of Cholet hospital contaminated with Aeromonas species was not the source of the cluster of hospital infections and only two patients were infected with clonally-related strains. RAPD using primer AP3 was simpler, cheaper, and quicker to perform than pulsed-field gel electrophoresis and is well suited for the epidemiological study of A. hydrophila isolates.

摘要

通过将任意引物聚合酶链反应(RAPD)与脉冲场凝胶电泳(PFGE)进行比较,评估了其作为一种分型技术的适用性,以对从一组医院获得性感染中分离出的嗜水气单胞菌菌株进行特征分析。将来自患者的5株分离株和来自供水系统的10株分离株与从患者和河流中分离出的10株流行病学上不相关的菌株进行比较。使用两种方法制备DNA,并选择了两种引物(AP3和AP5)。与煮沸法相比,提取DNA制备方法的鉴别力更好。使用AP3进行PCR对亲缘关系较近和较远的菌株进行鉴别,结果与PFGE一致:绍莱医院供水系统被气单胞菌属污染并非医院感染群的源头,只有两名患者感染了克隆相关菌株。使用引物AP3的RAPD比脉冲场凝胶电泳操作更简单、成本更低且速度更快,非常适合用于嗜水气单胞菌分离株的流行病学研究。

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