Schlegel Laurent, Grimont Francine, Grimont Patrick A D, Bouvet Anne
Centre National de Référence des Streptocoques, Service de Microbiologie, Hôtel Dieu, AP-HP Université Paris VI, Paris, France.
J Clin Microbiol. 2003 Feb;41(2):657-66. doi: 10.1128/JCM.41.2.657-666.2003.
Amplified ribosomal DNA restriction analysis (rrn-ARDRA) is based on PCR amplification and restriction of a fragment of rRNA genes including 16S and 23S genes and the intergenic spacer. rrn-ARDRA was evaluated for the identification of species within the genus Streptococcus: A total of 148 type and reference strains of pyogenic, oral, and group D streptococci were examined in order to construct a database for identification of streptococci. The amplified product was a single band approximately 4500 bp long. This amplicon was digested separately with three (HhaI, MboII, and Sau3A) restriction endonucleases. Respectively, 27, 26, and 28 major patterns were observed after HhaI, MboII, and Sau3A restrictions. Streptococcal strains belonging to different species had different patterns or different combination of patterns. An identification system based upon a combination of the three restriction patterns in a single database was then proposed. rrn-ARDRA was successfully applied to 11 clinical isolates whose identification to the species level was difficult to obtain by phenotypic analysis. Using a database of well-characterized strains, rrn-ARDRA is a powerful method for the identification of streptococcal isolates.
核糖体DNA限制性分析（rrn-ARDRA）基于对包括16S和23S基因以及基因间隔区在内的rRNA基因片段进行PCR扩增和限制性内切酶酶切。对rrn-ARDRA用于鉴定链球菌属内的菌种进行了评估：共检测了148株化脓性、口腔和D群链球菌的模式菌株和参考菌株，以便构建一个用于鉴定链球菌的数据库。扩增产物为一条约4500 bp长的单带。该扩增子分别用三种限制性内切酶（HhaI、MboII和Sau3A）进行酶切。HhaI、MboII和Sau3A酶切后分别观察到27、26和28种主要模式。属于不同菌种的链球菌菌株具有不同的模式或不同模式的组合。随后提出了一种基于在单个数据库中组合三种限制性模式的鉴定系统。rrn-ARDRA成功应用于11株临床分离株，这些分离株通过表型分析难以鉴定到菌种水平。利用一个特征明确的菌株数据库，rrn-ARDRA是鉴定链球菌分离株的一种有效方法。
