Hermans P W, Sluijter M, Hoogenboezem T, Heersma H, van Belkum A, de Groot R
Department of Pediatrics, University Hospital Rotterdam, Erasmus University Rotterdam, The Netherlands.
J Clin Microbiol. 1995 Jun;33(6):1606-12. doi: 10.1128/jcm.33.6.1606-1612.1995.
The aim of this study was to identify the strengths and weaknesses of five DNA fingerprint methods for epidemiological typing of Streptococcus pneumoniae. We investigated the usefulness of (i) ribotyping, (ii) BOX fingerprinting with the BOX repetitive sequence of S. pneumoniae as a DNA probe, (iii) PCR fingerprinting with a primer homologous to the enterobacterial repetitive intergenic consensus sequence, (iv) pulsed-field gel electrophoresis of large DNA fragments, and (v) restriction fragment end labeling to detect restriction fragment length polymorphism of small DNA fragments. Twenty-eight S. pneumoniae strains isolated from the blood and/or cerebrospinal fluid of 21 patients were analyzed. Genetic clustering among the 28 strains was independent of the DNA fingerprint technique used. However, the discriminatory power and the similarity values differed significantly among the individual techniques. BOX fingerprinting, pulsed-field gel electrophoresis, and restriction fragment end labeling provided the highest degree of discriminatory power. Furthermore, the ease with which computerized fingerprint analysis could be conducted also varied significantly among the techniques. Ribotyping, BOX fingerprinting, and restriction fragment end labeling were very suitable techniques for accurate computerized data analysis. Because of their high discriminatory potential and ease of accurate analysis, we conclude that BOX fingerprinting and restriction fragment end labeling are the most suitable techniques to type pneumococcal strains.
本研究的目的是确定用于肺炎链球菌流行病学分型的五种DNA指纹图谱方法的优缺点。我们研究了以下方法的实用性:(i)核糖体分型;(ii)以肺炎链球菌的BOX重复序列作为DNA探针的BOX指纹图谱分析;(iii)使用与肠杆菌重复基因间共有序列同源的引物进行PCR指纹图谱分析;(iv)大DNA片段的脉冲场凝胶电泳;以及(v)限制性片段末端标记以检测小DNA片段的限制性片段长度多态性。对从21名患者的血液和/或脑脊液中分离出的28株肺炎链球菌进行了分析。28株菌株之间的基因聚类与所使用的DNA指纹图谱技术无关。然而,不同技术之间的鉴别力和相似性值存在显著差异。BOX指纹图谱分析、脉冲场凝胶电泳和限制性片段末端标记提供了最高程度的鉴别力。此外,不同技术进行计算机化指纹分析的难易程度也有显著差异。核糖体分型、BOX指纹图谱分析和限制性片段末端标记是非常适合进行精确计算机化数据分析的技术。由于其高鉴别潜力和易于进行精确分析,我们得出结论,BOX指纹图谱分析和限制性片段末端标记是对肺炎球菌菌株进行分型的最合适技术。