Masuzawa T, Komikado T, Yanagihara Y
Department of Microbiology, School of Pharmaceutical Sciences, University of Shizuoka, Japan.
Clin Diagn Lab Immunol. 1997 Jan;4(1):60-3. doi: 10.1128/cdli.4.1.60-63.1997.
Restriction fragment length polymorphism (RFLP) analysis of the outer surface protein C (ospC) gene amplicon was used for rapid screening for genetic variability within Borrelia burgdorferi sensu stricto species and for detection of multiple borreliae in culture. Primers for the ospC gene amplified a fragment of about 600 bp from Borrelia cultures. After cleavage of the amplified products by MboI and DraI, eight different RFLP types were found among 13 B. burgdorferi sensu stricto strains from various sources and geographical areas, and three RFLP types were found among 10 representative isolates from skin biopsy specimens taken from patients residing on the eastern end of Long Island, New York (B. W. Berger, R. C. Johnson, C. Kodner, and L. Coleman, J. Clin. Microbiol. 30:359-361, 1992). These results suggested that the DNA organization of B. burgdorferi sensu stricto is heterogeneous not only globally but also within a localized geographical area and that the ospC-based typing approach could differentiate the B. burgdorferi sensu stricto. From the results obtained using mixed cultures of two different RFLP types of B. burgdorferi sensu stricto, contamination of at least 0.5% of different types of Borrelia cells in culture could be detected. This method could detect a multiple-B. burgdorferi sensu stricto infection in the bladders of mice experimentally infected with two different RFLP type strains. The present study showed that RFLP analysis of ospC-PCR products is a reliable method for epidemiological typing of B. burgdorferi sensu stricto and could be used for rapid detection of mixed Borrelia culture and multiple B. burgdorferi sensu stricto infections in animals, ticks, and patients.
采用外表面蛋白C（ospC）基因扩增片段的限制性片段长度多态性（RFLP）分析，对狭义伯氏疏螺旋体种内的遗传变异性进行快速筛查，并检测培养物中的多种疏螺旋体。ospC基因引物从伯氏疏螺旋体培养物中扩增出一段约600 bp的片段。用MboI和DraI酶切扩增产物后，在来自不同来源和地理区域的13株狭义伯氏疏螺旋体菌株中发现了8种不同的RFLP类型，在从纽约长岛东端居住的患者皮肤活检标本中分离出的10株代表性菌株中发现了3种RFLP类型（B. W. 伯杰、R. C. 约翰逊、C. 科德纳和L. 科尔曼，《临床微生物学杂志》30:359 - 361, 1992年）。这些结果表明，狭义伯氏疏螺旋体的DNA组织不仅在全球范围内，而且在局部地理区域内都是异质的，并且基于ospC的分型方法可以区分狭义伯氏疏螺旋体。从使用两种不同RFLP类型的狭义伯氏疏螺旋体混合培养物获得的结果来看，培养物中至少0.5%的不同类型疏螺旋体细胞污染能够被检测到。该方法能够检测出实验感染两种不同RFLP类型菌株的小鼠膀胱中的多重狭义伯氏疏螺旋体感染。本研究表明，ospC - PCR产物的RFLP分析是狭义伯氏疏螺旋体流行病学分型的可靠方法，可用于快速检测动物、蜱虫和患者中的混合疏螺旋体培养物以及多重狭义伯氏疏螺旋体感染。
