Wang G, van Dam A P, Spanjaard L, Dankert J
Department of Medical Microbiology, Academic Medical Center, University of Amsterdam, The Netherlands.
J Clin Microbiol. 1998 Mar;36(3):768-76. doi: 10.1128/JCM.36.3.768-776.1998.
To study whether pathogenic clusters of Borrelia burgdorferi sensu lato strains occur, we typed 136 isolates, cultured from specimens from patients (n = 49) with various clinical entities and from ticks (n = 83) or dogs (n = 4) from different geographic regions, by randomly amplified polymorphic DNA (RAPD) fingerprinting with four arbitrary primers. The RAPD patterns were reproducible up to the 95% similarity level as shown in duplicate experiments. In these experiments the purified DNAs prepared on different days, from different colonies, and after various passages were used as templates. With an intergroup difference of 55%, the 136 strains could be divided into seven genetic clusters. Six clusters comprised and corresponded to the established species B. burgdorferi sensu stricto (n = 23), Borrelia garinii (n = 39), Borrelia afzelii (n = 59), Borrelia japonica (n = 1), Borrelia valaisiana (n = 12), and genomic group DN127 (n = 1). One strain from a patient with erythema migrans (EM) did not belong to any of the species or genomic groups known up to now. The RAPD types of B. burgdorferi sensu stricto, B. garinii, and B. afzelii isolates, which may give rise to human Lyme borreliosis (LB), were associated with their geographic origins. A high degree of genetic diversity was observed among the 39 B. garinii strains, and six subgroups could be recognized. One of these comprised eight isolates from patients with disseminated LB only and no tick isolates. B. afzelii strains from patients with EM or acrodermatitis chronica atrophicans were not clustered in particular branches. Our study showed that RAPD analysis is a powerful tool for discriminating different Borrelia species as well as Borrelia isolates within species.
为研究伯氏疏螺旋体狭义菌株的致病菌群是否存在，我们用四种任意引物通过随机扩增多态性DNA（RAPD）指纹图谱技术，对136株分离株进行分型。这些分离株是从不同临床类型患者（49例）的标本以及来自不同地理区域的蜱（83只）或狗（4只）中培养得到的。重复实验表明，RAPD图谱在相似度达95%时仍具有可重复性。在这些实验中，不同日期、不同菌落以及经过多次传代后制备的纯化DNA均用作模板。由于组间差异为55%，这136株菌株可分为七个基因簇。六个簇包含并对应于已确定的物种狭义伯氏疏螺旋体（23株）、伽氏疏螺旋体（39株）、阿氏疏螺旋体（59株）、日本疏螺旋体（1株）、瓦氏疏螺旋体（12株）和基因组群DN127（1株）。一名患有游走性红斑（EM）患者的一株菌株不属于目前已知的任何物种或基因组群。可能引发人类莱姆病（LB）的狭义伯氏疏螺旋体、伽氏疏螺旋体和阿氏疏螺旋体分离株的RAPD类型与它们的地理来源相关。在39株伽氏疏螺旋体菌株中观察到高度的遗传多样性，可识别出六个亚组。其中一个亚组仅包含八株来自播散性LB患者的分离株，没有蜱分离株。患有EM或慢性萎缩性肢端皮炎患者的阿氏疏螺旋体菌株未聚集在特定分支中。我们的研究表明，RAPD分析是区分不同疏螺旋体物种以及物种内疏螺旋体分离株的有力工具。
