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用于奇异变形杆菌临床分离株特征鉴定的基于PCR的分子标记物比较

Comparison of PCR-based molecular markers for the characterization of Proteus mirabilis clinical isolates.

作者信息

Michelim Lessandra, Muller Gabriela, Zacaria Jucimar, Delamare Ana Paula Longaray, Costa Sérgio Olavo Pinto da, Echeverrigaray Sergio

机构信息

Institute of Biotechnology, University of Caxias do Sul, RS, Brazil.

出版信息

Braz J Infect Dis. 2008 Oct;12(5):423-9. doi: 10.1590/s1413-86702008000500014.

Abstract

Proteus mirabilis is one of the most important pathogens associated with complicated urinary tract infections (acute pyelonephritis, bladder infections, kidney stones) and bacteremia, affecting patients with anatomical abnormalities, immunodeficiency, and long-term urinary catheterization. For epidemiological purposes, various molecular typing methods, such as pulse-field gel electrophoresis (PFGE) or ribotyping, have been developed for this pathogen. However, these methods are labor intensive and time-consuming. We evaluated the discriminatory power of several PCR-based fingerprinting methods (RAPD, ISSR, ERIC-PCR, BOX-PCR and rep-PCR) for P. mirabilis clinical isolates. Typing patterns and clustering analysis indicated that RAPD, BOX-PCR and ERIC-PCR differentiated P. mirabilis strains from Escherichia coli, Hafnia alvei, and Morganella morganii. With the exception of rep-PCR, the methods gave medium to high discriminatory efficiency in P. mirabilis. In general, the results obtained with RAPD, BOX-PCR and ERIC-PCR were in good agreement. We concluded that a combination of ERIC-PCR and BOX-PCR results is a rapid and reliable alternative for discrimination among P. mirabilis clinical isolates, contributing to epidemiological studies.

摘要

奇异变形杆菌是与复杂性尿路感染(急性肾盂肾炎、膀胱感染、肾结石)和菌血症相关的最重要病原体之一,影响有解剖结构异常、免疫缺陷和长期留置导尿管的患者。出于流行病学目的,已针对这种病原体开发了多种分子分型方法,如脉冲场凝胶电泳(PFGE)或核糖体分型。然而,这些方法劳动强度大且耗时。我们评估了几种基于PCR的指纹图谱方法(随机扩增多态性DNA分析、简单序列重复区间PCR、肠杆菌基因间重复一致序列PCR、BOX-PCR和重复序列PCR)对奇异变形杆菌临床分离株的鉴别能力。分型模式和聚类分析表明,随机扩增多态性DNA分析、BOX-PCR和肠杆菌基因间重复一致序列PCR可区分奇异变形杆菌菌株与大肠杆菌、蜂房哈夫尼亚菌和摩根摩根菌。除重复序列PCR外,这些方法对奇异变形杆菌的鉴别效率为中等至高。总体而言,随机扩增多态性DNA分析、BOX-PCR和肠杆菌基因间重复一致序列PCR获得的结果高度一致。我们得出结论,肠杆菌基因间重复一致序列PCR和BOX-PCR结果的组合是区分奇异变形杆菌临床分离株的快速可靠替代方法,有助于流行病学研究。

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