Schwan T G, Simpson W J, Schrumpf M E, Karstens R H
Laboratory of Pathobiology, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, Montana 59840.
J Clin Microbiol. 1989 Aug;27(8):1734-8. doi: 10.1128/jcm.27.8.1734-1738.1989.
Fragments of plasmid DNA from Borrelia burgdorferi and B. hermsii were cloned and tested for specificity as hybridization probes to identify these two species of pathogenic spirochetes. Three fragments from the 49-kilobase-pair linear plasmid of B. burgdorferi were tested: a 500-base-pair (bp) HindIII fragment (probe 49A), a 445-bp PstI-HindIII fragment (probe 3G), and a 320-bp HindIII fragment (probe 16H). When hybridized to purified DNA or whole spirochetes, all of the probes distinguished B. burgdorferi from the other species examined, including B. hermsii, B. parkeri, B. turicatae, B. coriaceae, B. crocidurae, and B. anserina. Probe 49A was the most useful, however, hybridizing with all strains of B. burgdorferi originating from both North America and Europe while not cross-hybridizing with B. hermsii. A 790-bp HindIII fragment of B. hermsii DNA hybridized with DNA and whole spirochetes of this species and also with B. parkeri, confirming the close relatedness of these two species. These probes provide a new method of identifying these Borrelia species once the organisms have been grown in culture.
克隆了来自伯氏疏螺旋体和赫氏疏螺旋体的质粒DNA片段,并测试其作为杂交探针的特异性,以鉴定这两种致病性螺旋体。对来自伯氏疏螺旋体49千碱基对线性质粒的三个片段进行了测试:一个500碱基对(bp)的HindIII片段(探针49A)、一个445bp的PstI-HindIII片段(探针3G)和一个320bp的HindIII片段(探针16H)。当与纯化的DNA或完整的螺旋体杂交时,所有探针都能将伯氏疏螺旋体与其他检测的物种区分开来,包括赫氏疏螺旋体、帕克疏螺旋体、杜氏疏螺旋体、科氏疏螺旋体、麝鼩疏螺旋体和鹅疏螺旋体。然而,探针49A最有用,它能与来自北美和欧洲的所有伯氏疏螺旋体菌株杂交,而不与赫氏疏螺旋体交叉杂交。赫氏疏螺旋体DNA的一个790bp HindIII片段与该物种的DNA和完整螺旋体以及帕克疏螺旋体杂交,证实了这两个物种的密切相关性。一旦这些生物体在培养物中生长,这些探针提供了一种鉴定这些疏螺旋体物种的新方法。