Genotypic and phenotypic characterization of Borrelia burgdorferi isolated from ticks and small animals in Illinois.
Genotypic and phenotypic characterization of Borrelia burgdorferi isolated from ticks and small animals in Illinois.
复制标题
从伊利诺伊州蜱虫和小动物中分离出的伯氏疏螺旋体的基因型和表型特征。
DOI：
10.1128/jcm.33.9.2304-2315.1995
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发表时间：
1995
影响因子：
9.4
通讯作者：
Trenholme,GM
中科院分区：
文献类型：
--
作者：
Picken,RN;Cheng,Y;Han,D;Nelson,JA;Reddy,AG;Hayden,MK;Picken,MM;Strle,F;Bouseman,JK;Trenholme,GM
我们对来自伊利诺伊州北部（32 个分离株）和威斯康星州（1 个分离株）的 33 个伯氏疏螺旋体分离株进行了鉴定，代表了迄今为止调查的中西部分离株的最大系列。用于菌株分子分析的技术包括 (i) 使用物种特异性 PCR 引物进行基因种分型，(ii) 通过总基因组 DNA 的脉冲场凝胶电泳进行质粒分析，(iii) 通过 MluI 消化的基因组 DNA 的脉冲场凝胶电泳进行大限制性片段模式 (LRFP) 分析（J. Belfaiza、D. Postic、E. Bellenger、G. Baranton 和 I. Saint） Girons，J. Clin. 31：2873-2877，1993），（iv）总蛋白的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，（v）来自显示高水平表达的蛋白的高效液相色谱纯化的肽的微测序，（vi）蛋白的氨基酸组成分析，以及（vii）用人源多克隆抗血清对蛋白进行免疫学分析。包括五种参考菌株以及来自加利福尼亚州 (DN127) 和纽约州 (25015) 的两种非典型蜱分离株进行比较。所有伊利诺伊州和威斯康星州的分离株均使用基因种特异性 PCR 引物进行了严格型伯氏疏螺旋体的分类。发现分离株的质粒和蛋白质谱具有异质性。来自伊利诺伊州的一种分离株拥有两个大分子大小的质粒，而不是通常的 49 kb 质粒。对来自 33 个分离株和菌株 DN127 和 25015 的基因组 DNA 进行 MluI 消化所得的片段模式可分为 6 个不同的 LRFP，其中 5 个以前未曾描述过。菌株 25015 和来自伊利诺伊州的分离株 (CT39) 具有不寻常的 LRFP，这在其他伯氏疏螺旋体菌株中并不典型，表明它们可能代表伯氏疏螺旋体的第五种。 33 个分离株和菌株 DN127 和 25015 中的 5 个显示出分子量约为 22 kDa 的蛋白质的高水平表达。通过对单个肽进行微测序和总氨基酸组成分析对这些蛋白质进行研究表明，这 7 个菌株表达的 22-kDa 蛋白质是多态性 OspC 蛋白质。通过使用人源多克隆血清，可以在所有 33 个伊利诺伊州和威斯康星州分离株中检测到 OspC 的表达。
We have characterized 33 isolates of Borrelia burgdorferi from northern Illinois (32 isolates) and Wisconsin (1 isolate) representing the largest series of midwestern isolates investigated to date. The techniques used for molecular analysis of strains included (i) genospecies typing with species-specific PCR primers, (ii) plasmid profiling by pulsed-field gel electrophoresis of total genomic DNA, (iii) large-restriction-fragment pattern (LRFP) analysis by pulsed-field gel electrophoresis of MluI-digested genomic DNA (J. Belfaiza, D. Postic, E. Bellenger, G. Baranton, and I. Saint Girons, J. Clin. Microbiol. 31:2873-2877, 1993), (iv) sodium dodecyl sulfate-polyacrylamide gel electrophoresis of total proteins, (v) microsequencing of high-performance liquid chromatography-purified peptides derived from proteins showing high levels of expression, (vi) amino acid composition analysis of proteins, and (vii) immunological analysis of proteins with a polyclonal antiserum of human origin. Five reference strains as well as two atypical tick isolates from California (DN127) and New York (25015) were included for comparison. All of the Illinois and Wisconsin isolates were typed as B. burgdorferi sensu stricto with genospecies-specific PCR primers. The isolates were found to be heterogeneous with regard to their plasmid and protein profiles. One isolate from Illinois possessed two large-molecular-size plasmids instead of the usual 49-kb plasmid. Fragment patterns resulting from MluI digestion of genomic DNA from the 33 isolates and strains DN127 and 25015 were separable into six distinct LRFPs, five of which have not previously been described. Strain 25015 and an isolate from Illinois (CT39) shared an unusual LRFP that is not typical of other B. burgdorferi sensu stricto strains, suggesting that they may represent a fifth species of B. burgdorferi sensu lato. Five of the 33 isolates and strains DN127 and 25015 showed high-level expression of proteins with molecular masses of approximately 22 kDa. Investigation of these proteins by microsequencing of individual peptides and total amino acid composition analysis indicated that the 22-kDa proteins expressed by the seven strains were polymorphic OspC proteins. By using a polyclonal serum of human origin, expression of OspC could be detected in all 33 Illinois and Wisconsin isolates.