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用于不动杆菌基因组种鉴定及鲍曼不动杆菌分型的扩增核糖体DNA限制性分析、随机扩增多态性DNA分析和扩增片段长度多态性指纹图谱比较

Comparison of amplified ribosomal DNA restriction analysis, random amplified polymorphic DNA analysis, and amplified fragment length polymorphism fingerprinting for identification of Acinetobacter genomic species and typing of Acinetobacter baumannii.

作者信息

Koeleman J G, Stoof J, Biesmans D J, Savelkoul P H, Vandenbroucke-Grauls C M

机构信息

Department of Clinical Microbiology and Infection Control, University Hospital Vrije Universiteit, 1007 MB Amsterdam, The Netherlands.

出版信息

J Clin Microbiol. 1998 Sep;36(9):2522-9. doi: 10.1128/JCM.36.9.2522-2529.1998.

Abstract

Thirty-one strains of Acinetobacter species, including type strains of the 18 genomic species and 13 clinical isolates, were compared by amplified ribosomal DNA restriction analysis (ARDRA), random amplified polymorphic DNA analysis (RAPD), and amplified fragment length polymorphism (AFLP) fingerprinting. ARDRA, performed with five different enzymes, showed low discriminatory power for differentiating Acinetobacter at the species and strain level. The standardized commercially available RAPD kit clearly enabled the discrimination of all Acinetobacter genomic species but showed great polymorphism between isolates of Acinetobacter baumannii. AFLP fingerprinting with radioactively as well as fluorescently labelled primers showed high discriminatory power for the identification of 18 Acinetobacter genomic species and typing of 13 clinical Acinetobacter isolates. Compared to radioactive AFLP, fluorescent AFLP was technically fast and simple to perform, and it permitted analysis with an automated DNA sequencer. Fluorescent AFLP seems particularly well suited for studying the epidemiology of nosocomial infections and outbreaks caused by Acinetobacter species.

摘要

通过扩增核糖体DNA限制性分析(ARDRA)、随机扩增多态性DNA分析(RAPD)和扩增片段长度多态性(AFLP)指纹分析,对31株不动杆菌属菌株进行了比较,其中包括18个基因组种的模式菌株和13株临床分离株。使用5种不同酶进行的ARDRA在区分不动杆菌属的种和菌株水平上显示出较低的鉴别能力。标准化的市售RAPD试剂盒能够清楚地区分所有不动杆菌属基因组种,但鲍曼不动杆菌分离株之间表现出很大的多态性。使用放射性和荧光标记引物的AFLP指纹分析在鉴定18个不动杆菌属基因组种和对13株临床不动杆菌分离株进行分型方面显示出高鉴别能力。与放射性AFLP相比,荧光AFLP在技术上快速且操作简单,并且允许使用自动DNA测序仪进行分析。荧光AFLP似乎特别适合研究由不动杆菌属引起的医院感染和暴发的流行病学。

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