Misonne M C, Hoet P P
Microbial Pathogenesis Unit, Catholic University of Louvain Medical School, International Institute of Cellular and Molecular Pathology, Brussels, Belgium.
J Clin Microbiol. 1998 Jan;36(1):269-72. doi: 10.1128/JCM.36.1.269-272.1998.
Species-specific sequences were shown to be carried by plasmids of the three main species of Borrelia burgdorferi sensu lato involved in Lyme disease. Libraries of the 16-, 33-, and 25-kb plasmids of B. burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii, respectively, were then built and used to isolate species-specific sequences. After sequencing of the cloned inserts, three sets of primers were designed. They were shown to determine species-specific PCR amplification products. The sensitivities of the PCR assay with these primers were 100 spirochetes for B. burgdorferi sensu stricto and 1,000 spirochetes for B. garinii and B. afzelii. The usefulness of these primers for the identification of species in biological samples (tick, serum, and cerebrospinal fluid samples) was ascertained.
研究表明，莱姆病相关的伯氏疏螺旋体狭义种的三个主要菌种的质粒携带种特异性序列。随后分别构建了狭义伯氏疏螺旋体、伽氏疏螺旋体和阿氏疏螺旋体的16kb、33kb和25kb质粒文库，并用于分离种特异性序列。对克隆插入片段进行测序后，设计了三组引物。结果表明，这些引物可用于确定种特异性PCR扩增产物。使用这些引物进行PCR检测的灵敏度，对于狭义伯氏疏螺旋体为100个螺旋体，对于伽氏疏螺旋体和阿氏疏螺旋体为1000个螺旋体。已确定这些引物在生物样品（蜱、血清和脑脊液样品）中鉴定菌种的实用性。
