Güner Ece S, Hashimoto Naoya, Kadosaka Teruki, Imai Yasuyuki, Masuzawa Toshiyuki
Medical School, Yeditepe University, Istanbul, Turkey.
Department of Microbiology, and COE Program in the 21st Century, University of Shizuoka School of Pharmaceutical Sciences, Shizuoka, 422-8526, Japan.
Microbiology (Reading). 2003 Sep;149(Pt 9):2539-2544. doi: 10.1099/mic.0.26464-0.
A novel, fast-growing spirochaete was isolated from the hard tick Hyalomma aegyptium (family Ixodidae, subfamily Metastriata) using Barbour-Stoenner-Kelly (BSK) II medium. Tick samples were taken during the summer of 2000 from the Istanbul area in northwestern Turkey. Sixty-seven of 153 adults (44%) and 72 of 185 nymphs (39%) were infected with the novel spirochaete, whereas none of the 20 larvae examined were infected. The optimal growth temperature of the spirochaete in BSK II medium was 34-37 degrees C, and it could grow at 39 degrees C. Doubling times at 34 and 37 degrees C were 5.3 and 5.1 h, respectively. Six pure cultures of the spirochaete were obtained and characterized by microscopic observation, sequence analysis of the flagellin gene (flaB), SDS-PAGE and Western blotting. The spirochaete was morphologically similar to those of the genus Borrelia and contained a 41 kDa protein reactive with mAb H9724 specific to the flagellin of a Borrelia species. Polyclonal antibody raised to this spirochaete reacted with several antigen bands, whereas no bands were detected with Borrelia burgdorferi, Borrelia hermsii, Borrelia turicatae and Borrelia parkeri. The flaB sequences of the six isolates showed high similarity, with sequence similarity values ranging from 99.2 to 100%; however, the similarity of the isolates' flaB sequences to those of the Lyme-disease-related Borrelia and relapsing-fever-associated Borrelia species was less than 90%. These findings suggest that the unique spirochaete is a member of the genus Borrelia, and differs from previously described Borrelia species.
利用巴伯-斯托纳-凯利（BSK）II培养基，从硬蜱埃及璃眼蜱（蜱科，后沟蜱亚科）中分离出一种新型的、生长迅速的螺旋体。2000年夏季，在土耳其西北部的伊斯坦布尔地区采集蜱样本。153只成虫中有67只（44%）、185只若虫中有72只（39%）感染了这种新型螺旋体，而所检查的20只幼虫均未感染。该螺旋体在BSK II培养基中的最佳生长温度为34 - 37摄氏度，在39摄氏度时也能生长。在34摄氏度和37摄氏度下的倍增时间分别为5.3小时和5.1小时。获得了该螺旋体的6个纯培养物，并通过显微镜观察、鞭毛蛋白基因（flaB）序列分析、SDS - PAGE和蛋白质免疫印迹法进行了鉴定。该螺旋体在形态上与疏螺旋体属的螺旋体相似，并且含有一种41 kDa的蛋白质，可与针对疏螺旋体属某一物种鞭毛蛋白的单克隆抗体H9724发生反应。针对这种螺旋体产生的多克隆抗体与几条抗原带发生反应，而与伯氏疏螺旋体、赫氏疏螺旋体、土耳其疏螺旋体和帕克疏螺旋体均未检测到条带。6个分离株的flaB序列显示出高度相似性，序列相似性值在99.2%至100%之间；然而，这些分离株的flaB序列与莱姆病相关疏螺旋体和回归热相关疏螺旋体物种的序列相似性小于90%。这些发现表明，这种独特的螺旋体是疏螺旋体属成员，且与先前描述的疏螺旋体物种不同。
