Department of Genetics, University of Szczecin, 71-065 Szczecin, Poland.
J Med Microbiol. 2010 Mar;59(Pt 3):309-314. doi: 10.1099/jmm.0.013508-0. Epub 2009 Dec 10.
Borrelia burgdorferi sensu lato, carried by Ixodes ticks, is one of the most significant human pathogens, causing Lyme disease. As there is no standardized PCR method for detection and identification of spirochaete DNA, we carried out a comparative analysis using a set of complementary primers for three regions in the genomic DNA of these bacteria (genes fla and rrs and the non-coding rrs-rrlA region). DNA extracted from 579 Ixodes ricinus ticks was subjected to nested PCR. DNA of the examined spirochaetes was detected in 43 (7.4 %) lysates when the fla gene was used as a molecular marker, in 7 (1.2 %) lysates when using primers complementary to the rrs gene, and in 12 (2.1 %) lysates using primers complementary to the non-coding rrs-rrlA sequence. RFLP analysis based on the fla gene helped identify species from the B. burgdorferi sensu lato complex (B. burgdorferi sensu stricto, Borrelia afzelii, Borrelia garinii, Borrelia valaisiana), detect co-infections, and also identify Borrelia miyamotoi. Therefore, the fla gene is the most sensitive and specific molecular marker for the detection and identification of Borrelia spirochaetes in I. ricinus.
伯氏疏螺旋体（Borrelia burgdorferi sensu lato）通过硬蜱传播，是最重要的人类病原体之一，可引起莱姆病。由于目前尚无用于检测和鉴定螺旋体 DNA 的标准化 PCR 方法，我们使用针对这些细菌基因组 DNA 中三个区域的一组互补引物进行了比较分析（fla 和 rrs 基因以及非编码 rrs-rrlA 区）。对 579 只硬蜱的 DNA 进行了巢式 PCR 检测。当使用 fla 基因为分子标记时，在 43 个（7.4%）裂解物中检测到被检螺旋体的 DNA，当使用与 rrs 基因互补的引物时，在 7 个（1.2%）裂解物中检测到，当使用与非编码 rrs-rrlA 序列互补的引物时，在 12 个（2.1%）裂解物中检测到。基于 fla 基因的 RFLP 分析有助于鉴定伯氏疏螺旋体复合体中的种（伯氏疏螺旋体、阿费尔约森螺旋体、伽氏疏螺旋体、瓦氏疏螺旋体），检测共感染，还可鉴定伯氏疏螺旋体。因此，fla 基因是检测和鉴定硬蜱中伯氏疏螺旋体的最敏感和特异的分子标记。
