Schwan T G, Schrumpf M E, Karstens R H, Clover J R, Wong J, Daugherty M, Struthers M, Rosa P A
Arthropod-Borne Diseases Section, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, Montana 59840.
J Clin Microbiol. 1993 Dec;31(12):3096-108. doi: 10.1128/jcm.31.12.3096-3108.1993.
Previous studies describing the occurrence and molecular characteristics of Lyme disease spirochetes, Borrelia burgdorferi, from California have been restricted primarily to isolates obtained from the north coastal region of this large and ecologically diverse state. Our objective was to look for and examine B. burdorferi organisms isolated from Ixodes pacificus ticks collected from numerous regions spanning most parts of California where this tick is found. Thirty-one isolates of B. burgdorferi were examined from individual or pooled I. pacificus ticks collected from 25 counties throughout the state. One isolate was obtained from ticks collected at Wawona Campground in Yosemite National Park, documenting the occurrence of the Lyme disease spirochete in an area of intensive human recreational use. One isolate from an Ixodes neotomae tick from an additional county was also examined. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblot analysis, agarose gel electrophoresis, Southern blot analysis, and the polymerase chain reaction were used to examine the molecular and genetic determinants of these uncloned, low-passage-number isolates. All of the isolates were identified as B. burgdorferi by their protein profiles and reactivities with monoclonal and polyclonal antibodies, and all the isolates were typed by the polymerase chain reaction as North American-type spirochetes (B. burgdorferi sensu stricto). Although products of the ospAB locus were identified in protein analyses in all of the isolates, several isolates contained deleted forms of this locus that would result in the expression of chimeric OspA-OspB proteins. The analysis of OspC demonstrated that this protein was widely conserved among the isolates but was also quite variable in its molecular mass and the amount of it that was expressed.
先前描述来自加利福尼亚州的莱姆病螺旋体——伯氏疏螺旋体的发生情况及分子特征的研究,主要局限于从这个地域广阔且生态多样的州的北部沿海地区分离得到的菌株。我们的目标是寻找并检测从太平洋硬蜱中分离出的伯氏疏螺旋体,这些蜱虫采自加利福尼亚州大部分地区发现该蜱虫的众多区域。从全州25个县采集的单个或混合的太平洋硬蜱中检测了31株伯氏疏螺旋体。从优胜美地国家公园瓦沃纳露营地采集的蜱虫中获得了一株菌株,证明莱姆病螺旋体在人类密集休闲活动区域存在。还检测了来自另一个县的一只新英格兰硬蜱的一株菌株。使用十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳、免疫印迹分析、琼脂糖凝胶电泳、Southern印迹分析和聚合酶链反应来检测这些未克隆的低传代菌株的分子和遗传决定因素。所有菌株通过其蛋白质谱以及与单克隆和多克隆抗体的反应性被鉴定为伯氏疏螺旋体,并且所有菌株通过聚合酶链反应被分型为北美型螺旋体(狭义伯氏疏螺旋体)。尽管在所有菌株的蛋白质分析中都鉴定出了ospAB基因座的产物,但有几株菌株含有该基因座的缺失形式,这将导致嵌合OspA - OspB蛋白的表达。对OspC的分析表明,该蛋白在菌株中广泛保守,但其分子量和表达量也有很大差异。