随机扩增多态性DNA的自动激光荧光分析：一种调查鲍曼不动杆菌医院内传播的快速方法。

Automated laser fluorescence analysis of randomly amplified polymorphic DNA: a rapid method for investigating nosocomial transmission of Acinetobacter baumannii.

作者信息

Grundmann H, Schneider C, Tichy H V, Simon R, Klare I, Hartung D, Daschner F D

机构信息

Institut für Umweltmedizin und Krankenhaushygiene, Universitätsklinik Freiburg, Germany.

出版信息

J Med Microbiol. 1995 Dec;43(6):446-51. doi: 10.1099/00222615-43-6-446.

DOI:10.1099/00222615-43-6-446

PMID:7473679

Abstract

A rapid method for genotyping Acinetobacter baumannii based on PCR-fingerprinting with fluorescent primers was evaluated. Automated laser fluorescence analysis (ALFA) enabled on-line generation of high resolution DNA-fingerprints during polyacrylamide gel electrophoresis of randomly amplified polymorphic DNA (RAPD) products. The results were in concordance with macro-restriction fragment patterns produced by pulse-field gel electrophoresis (PFGE) of ApaI digests of chromosomal DNA. RAPD-ALFA was able to identify homologous strains suggestive of horizontal transmission in < 8 h after colonies were obtained on solid media, whereas PFGE analysis took c. 90 h. Speed and digitised data format renders RAPD-ALFA attractive for routine in-house epidemiological screening of isolates from intensive care and other hospital units.

摘要

对一种基于使用荧光引物进行PCR指纹分析的鲍曼不动杆菌基因分型快速方法进行了评估。自动激光荧光分析（ALFA）能够在随机扩增多态性DNA（RAPD）产物的聚丙烯酰胺凝胶电泳过程中在线生成高分辨率DNA指纹。结果与染色体DNA的ApaI酶切产物经脉冲场凝胶电泳（PFGE）产生的宏观限制性片段模式一致。RAPD-ALFA能够在固体培养基上获得菌落后不到8小时内鉴定出提示水平传播的同源菌株，而PFGE分析大约需要90小时。速度和数字化数据格式使RAPD-ALFA对于重症监护病房和其他医院科室分离株的常规内部流行病学筛查具有吸引力。