Ružić-Sabljić Eva, Maraspin Vera, Stupica Daša, Rojko Tereza, Bogovič Petra, Strle Franc, Cerar Tjaša
Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia.
Department of Infectious Diseases, University Medical Centre Ljubljana, Ljubljana, Slovenia.
PLoS One. 2017 Feb 7;12(2):e0171622. doi: 10.1371/journal.pone.0171622. eCollection 2017.
The isolation of B. burgdorferi sensu lato requires the use of complex cultivation media. The aim of the study was to compare the usefulness of BSK-H (a commercial medium produced by HiMedia, India) and MKP medium. MKP and BSK-H media were prepared in accordance with the relevant protocols. Borrelia strains and skin culture biopsies were simultaneously inoculated into both media, incubated and checked for growth. Borrelial growth characteristics, isolation rates and characteristics of the isolated borreliae were analysed and compared. Initially, numbers of spirochaetes were higher in BSK-H than in MKP; however, in comparison with MKP, the strains subcultured in BSK-H medium were more frequently irregular, thin and non-motile, and rapidly died. In addition, the borrelial isolation rate from erythema migrans skin samples was higher in MKP than in BSK-H medium (108/171, 63.2% versus 70/171, 40.9%; p<0.0001). The far most frequently isolated species was Borrelia afzelii (92.9% and 97.2% strains isolated from BSK-H and MKP, respectively). Comparison of strains cultured from individual patients in both media showed differences in plasmid contents in 9/46 (19.6%) strain pairs, and protein profiles differed in 30/43 (69.8%) strain pairs, most often in the expression of OspC (in 27/28 patients OspC was expressed only in strains growing in MKP). BSK-H medium supports the growth of borrelial strains but MKP is superior with regard to the isolation rate, morphology and motility of strains. BSK-H medium supports fast initial growth of borreliae but this is followed by rapid deformation and death of the spirochaetes.
莱姆病疏螺旋体狭义种的分离需要使用复杂的培养基。本研究的目的是比较BSK-H（一种由印度HiMedia公司生产的商业培养基）和MKP培养基的实用性。MKP和BSK-H培养基均按照相关方案制备。将疏螺旋体菌株和皮肤培养活检组织同时接种到两种培养基中，进行培养并检查生长情况。分析并比较疏螺旋体的生长特性、分离率以及分离出的疏螺旋体的特征。最初，BSK-H培养基中螺旋体的数量高于MKP培养基；然而，与MKP相比，在BSK-H培养基中继代培养的菌株更常出现不规则、纤细且无运动性的情况，并迅速死亡。此外，MKP培养基中来自游走性红斑皮肤样本的疏螺旋体分离率高于BSK-H培养基（分别为108/171，63.2% 对70/171，40.9%；p<0.0001）。最常分离出的菌种是阿氏疏螺旋体（分别从BSK-H和MKP中分离出的菌株占92.9%和97.2%）。对在两种培养基中培养的来自个体患者的菌株进行比较，发现9/46（19.6%）的菌株对在质粒含量上存在差异，30/43（69.8%）的菌株对在蛋白质谱上存在差异，最常见的是OspC表达的差异（在27/28例患者中，OspC仅在MKP培养基中生长的菌株中表达）。BSK-H培养基支持疏螺旋体菌株的生长，但MKP在菌株的分离率、形态和运动性方面更具优势。BSK-H培养基支持疏螺旋体的快速初始生长，但随后螺旋体会迅速变形并死亡。
