Mahenthiralingam E, Campbell M E, Henry D A, Speert D P
Department of Paediatrics, University of British Columbia, Vancouver, Canada.
J Clin Microbiol. 1996 Dec;34(12):2914-20. doi: 10.1128/jcm.34.12.2914-2920.1996.
We fingerprinted a collection of 627 Burkholderia cepacia isolates from 255 patients with cystic fibrosis (CF) and 43 patients without CF and from the environment, by a PCR-based randomly amplified polymorphic DNA (RAPD) method with primers selected for their ability to produce discriminatory polymorphisms. The RAPD typing method was found to be reproducible and discriminatory, more sensitive than PCR ribotyping, and able to group epidemiologically related B. cepacia strains previously typed by both pulsed-field gel electrophoresis and conventional ribotyping. Seven strain types infecting multiple CF patients were found at several different CF treatment centers in Canada, the United States, the United Kingdom, France, and Australia, indicating the presence of epidemic strain types. Most CF patients were each colonized with a single strain type, and several patients harbored the same strain type for 5 or more years. B. cepacia isolates recovered from other clinical sources (44 isolates examined) and from the environment (58 isolates examined) possessed RAPD fingerprints that were generally distinct from CF-associated strain types (525 isolates examined). RAPD is a versatile fingerprinting method for studying the epidemiology of B. cepacia.
我们采用基于聚合酶链反应(PCR)的随机扩增多态性DNA(RAPD)方法,使用因其能产生鉴别性多态性而选择的引物,对来自255例囊性纤维化(CF)患者、43例非CF患者以及环境的627株洋葱伯克霍尔德菌分离株进行了指纹图谱分析。结果发现,RAPD分型方法具有可重复性和鉴别性,比PCR核糖体分型更敏感,并且能够对先前通过脉冲场凝胶电泳和传统核糖体分型鉴定的具有流行病学相关性的洋葱伯克霍尔德菌菌株进行分组。在加拿大、美国、英国、法国和澳大利亚的几个不同CF治疗中心发现了七种感染多名CF患者的菌株类型,这表明存在流行菌株类型。大多数CF患者各自被单一菌株类型定植,并且有几名患者携带相同菌株类型达5年或更长时间。从其他临床来源(检查了44株分离株)和环境(检查了58株分离株)中回收的洋葱伯克霍尔德菌分离株所具有的RAPD指纹图谱通常与CF相关菌株类型(检查了525株分离株)不同。RAPD是一种用于研究洋葱伯克霍尔德菌流行病学的通用指纹图谱方法。