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通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和单克隆抗体对芬兰莱姆病螺旋体狭义种分离株进行鉴定。

Characterization of Finnish Borrelia burgdorferi sensu lato isolates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and with monoclonal antibodies.

作者信息

Tuomi J, Rantamäki L K, Tanskanen R, Junttila J

机构信息

Department of Microbiology and Epizootology, College of Veterinary Medicine, Helsinki, Finland.

出版信息

J Clin Microbiol. 1995 Aug;33(8):1989-96. doi: 10.1128/jcm.33.8.1989-1996.1995.

Abstract

Thirty-seven Borrelia burgdorferi strains, isolated in 1992 from Ixodes ricinus in Finland, were investigated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and by immunoblotting and indirect immunofluorescence assay (IFA) with five to nine monoclonal antibodies (MAbs). By SDS-PAGE results and reactivities to MAbs H3TS, J 8.3, I 17.3, and D6, the 37 isolates were assigned to the species B. burgdorferi sensu stricto (n = 7), Borrelia afzelii (n = 17), or Borrelia garinii (n = 13). Twenty more isolates examined only by IFA and with part of the MAbs were distributed as follows: 9 B. burgdorferi sensu stricto and 11 other species. Among 16 of 37 isolates displaying a SDS-PAGE patterns considered typical of that of B. garinii, 3 were negative by the test with MAb D6; the rest were positive. The three MAb D6-negative isolates reacted with MAb J 8.3 but not with MAb I 17.3. It is suggested that these isolates of a previously undescribed type represent atypical B. afzelii strains deficient in the expression of OspB proteins. The misleading species designation by the SDS-PAGE result is described. The IFA results were generally consistent with those obtained by immunoblotting. The exception was for 3 of 29 isolates that were positive with MAb H5332 by immunoblotting but that were IFA negative. In the present material of 57 strains, all 16 B. burgdorferi sensu stricto isolates originated from the Aland Islands. B. afzelii and B. garinii were isolated from all three regions where ticks were collected. The distributive difference seems to offer a basis for comparative clinico-epidemiological studies of Lyme borreliosis.

摘要

1992年从芬兰蓖麻硬蜱中分离出的37株伯氏疏螺旋体菌株,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)、免疫印迹法以及使用5至9种单克隆抗体(MAb)的间接免疫荧光试验(IFA)进行了研究。根据SDS-PAGE结果以及对单克隆抗体H3TS、J 8.3、I 17.3和D6的反应性,将这37株分离株归为狭义伯氏疏螺旋体种(n = 7)、阿氏疏螺旋体(n = 17)或伽氏疏螺旋体(n = 13)。另外20株仅通过IFA并用部分单克隆抗体检测的分离株分布如下:9株狭义伯氏疏螺旋体和11株其他种。在37株显示出被认为是伽氏疏螺旋体典型SDS-PAGE图谱的分离株中,有16株,其中3株用单克隆抗体D6检测为阴性;其余为阳性。这3株单克隆抗体D6阴性的分离株与单克隆抗体J 8.3反应,但不与单克隆抗体I 17.3反应。提示这些先前未描述类型的分离株代表了阿氏疏螺旋体的非典型菌株,其OspB蛋白表达缺失。描述了SDS-PAGE结果导致的误导性种属命名。IFA结果总体上与免疫印迹法获得的结果一致。例外情况是29株分离株中有3株通过免疫印迹法用单克隆抗体H5332检测为阳性,但IFA检测为阴性。在目前的57株菌株材料中,所有16株狭义伯氏疏螺旋体分离株均来自奥兰群岛。阿氏疏螺旋体和伽氏疏螺旋体是从采集蜱的所有三个地区分离出来的。这种分布差异似乎为莱姆病螺旋体病的比较临床流行病学研究提供了基础。

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