Cerar T, Ruzić-Sabljić E, Glinsek U, Zore A, Strle F
Institute of Microbiology and Immunology, Faculty of Medicin, University of Ljubljana, Slovenia.
Clin Microbiol Infect. 2008 Jul;14(7):653-8. doi: 10.1111/j.1469-0691.2008.02013.x.
The sensitivities of two PCR assays and culture were compared for the detection of Borrelia spp. in skin specimens of 150 patients with typical erythema migrans. In addition, the accuracy of the methods for the identification of Borrelia spp. was compared by analysing culture isolates and material obtained directly from skin biopsy specimens. Borrelia burgdorferi sensu lato was isolated from 73 (49%) of 150 skin biopsy specimens. Using a nested PCR targeting the rrf-rrl region and a PCR targeting the flagellin gene, 107 (71%) and 36 (24%) specimens, respectively, were positive. With both PCRs, positive results were more frequent with culture-positive samples (67/73 (92%) and 24/73 (33%) for the nested and flagellin PCRs, respectively) than with culture-negative samples (40/77 (52%) and 12/77 (16%) for nested and flagellin PCR, respectively). Pulsed-field gel electrophoresis after MluI restriction identified 69/73 (95%) isolates, of which 58/69 (84%) were Borrelia afzelii and 11/69 (16%) were Borrelia garinii. After MseI restriction of PCR products amplified from the intergenic rrf-rrl region, B. afzelii was identified in 73/107 (68%) samples, B. garinii in 22/107 (21%) samples, and both species in 11/107 (10%) samples. The corresponding results for culture-positive specimens were 41/69 (59%), 14/69 (20%), and 7/69 (10%). Comparison of the results for specimens positive according to both approaches revealed complete uniformity in 80% of the cases. Overall, nested PCR was the most sensitive method for the demonstration of Borrelia spp. in erythema migrans skin lesions, followed by culture and PCR targeting the flagellin gene. The congruence of identification results obtained by analyzing culture isolates and material obtained directly from skin biopsies was relatively high but incomplete.
比较了两种聚合酶链反应（PCR）检测方法和培养法对150例典型游走性红斑患者皮肤标本中疏螺旋体属的检测灵敏度。此外，通过分析培养分离株和直接从皮肤活检标本中获取的材料，比较了鉴定疏螺旋体属方法的准确性。从150份皮肤活检标本中的73份（49%）分离出伯氏疏螺旋体狭义种。使用靶向rrf-rrl区域的巢式PCR和靶向鞭毛蛋白基因的PCR，分别有107份（71%）和36份（24%）标本呈阳性。两种PCR方法中，培养阳性样本的阳性结果比培养阴性样本更常见（巢式PCR为67/73（92%），鞭毛蛋白PCR为24/73（33%）；培养阴性样本中，巢式PCR为40/77（52%），鞭毛蛋白PCR为12/77（16%））。MluI酶切后的脉冲场凝胶电泳鉴定出69/73（95%）的分离株，其中58/69（84%）为阿氏疏螺旋体，11/69（16%）为伽氏疏螺旋体。对从基因间rrf-rrl区域扩增的PCR产物进行MseI酶切后，在73/107（68%）的样本中鉴定出阿氏疏螺旋体，22/107（21%）的样本中鉴定出伽氏疏螺旋体，11/107（10%）的样本中同时鉴定出这两种菌株。培养阳性标本的相应结果分别为41/69（59%）、14/69（20%）和7/69（10%）。两种方法均呈阳性的标本结果比较显示80%的病例完全一致。总体而言，巢式PCR是检测游走性红斑皮肤病变中疏螺旋体属最敏感的方法，其次是培养法和靶向鞭毛蛋白基因的PCR。通过分析培养分离株和直接从皮肤活检中获取的材料所获得的鉴定结果一致性较高但不完全一致。
