Sakallah S, Pasculle W, Lanning R, McDevitt D, Cooper D
Department of Pathology, University of Pittsburgh Medical Center, PA 15261, USA.
Mol Cell Probes. 1995 Dec;9(6):405-14. doi: 10.1006/mcpr.1995.0063.
A new method to identify clonal strains of pathogenic bacteria has been developed recently in this laboratory. The method utilizes degenerate random amplified polymorphic DNA primers (D-RAPD) to amplify random fragments in crude bacterial lysates, generating reproducible DNA banding profiles or fingerprints. We use this method to type outbreak and non-outbreak isolates of Legionella pneumophila serogroup 1 from four hospitals near to, and affiliated with the University of Pittsburgh Medical Center. Patient isolates from a large outbreak, and nearly half of the contemporaneous environmental isolates showed the same DNA profile. Other isolates derived from non-outbreak patients showed easily distinguishable profiles. Other Legionella isolates collected between 1984 and 1994 were also analysed by this method. Our studies demonstrate that four strains were common among patient and environmental isolates at the four hospitals. These strains were also found to be different from a limited number of isolates from outside the Pittsburgh area. Because of its speed, simplicity and powerful discriminating ability, we believe that the D-RAPD approach provides epidemiologists and hospital infection control teams with a powerful tool in their efforts in analysing and terminating infection outbreaks.
本实验室最近开发了一种鉴定病原菌克隆菌株的新方法。该方法利用简并随机扩增多态性DNA引物(D-RAPD)扩增细菌粗裂解物中的随机片段,生成可重复的DNA条带图谱或指纹图谱。我们使用这种方法对匹兹堡大学医学中心附近四家医院的1型嗜肺军团菌暴发株和非暴发株进行分型。来自一次大型暴发的患者分离株以及近一半同期环境分离株显示出相同的DNA图谱。来自非暴发患者的其他分离株显示出易于区分的图谱。还使用该方法分析了1984年至1994年间收集的其他军团菌分离株。我们的研究表明,这四家医院的患者和环境分离株中有四种菌株是常见的。还发现这些菌株与匹兹堡地区以外数量有限的分离株不同。由于其速度快、操作简单且鉴别能力强,我们认为D-RAPD方法为流行病学家和医院感染控制团队在分析和终止感染暴发的工作中提供了一个强大的工具。