Bruneau S, de Montclos H, Drouet E, Denoyel G A
Institut Pasteur de Lyon, France.
J Clin Microbiol. 1994 Dec;32(12):2953-8. doi: 10.1128/jcm.32.12.2953-2958.1994.
The use of rRNA gene restriction fragment length polymorphism analysis (ribotyping) to subtype Streptococcus pyogenes strains was investigated. Sixty-eight S. pyogenes strains, including 17 reference strains and 51 isolates from blood, acute or chronic pharyngitis, and food-borne outbreaks, were characterized by determination of both their rDNA restriction fragment length polymorphism profiles and their serotypes (T and M). Total DNA was cleaved with five selected restriction enzymes and then probed with a digoxigenin-labeled stretch of 1,063 bp hybridizing with 16S rRNA genes. Fifteen and nine distinct patterns were generated with SacI and XhoI, respectively, and five patterns were generated with each of the three additional restriction enzymes. With the combination SacI-XhoI, a total of 21 distinct ribotypes were obtained among the 68 isolates. This number was not increased by the results obtained with the other restriction enzymes. All strains tested were typeable. All isolates from each food-borne outbreak belonged to the same ribotype, and all isolates (pre- and posttreatment) from each child with chronic pharyngitis also belonged to the same ribotype, suggesting antibiotic treatment failures. A discriminatory index was calculated for the 47 isolates which were epidemiologically unrelated, using the Hunter-Gaston formula. This index reached 0.955 when the combination SacI-XhoI was used, showing the good discriminatory power of this typing method. Therefore, ribotyping proved to be a molecular method of interest to subtype S. pyogenes. Moreover, there was some correlation between ribotyping and serotyping, as several ribotypes were related to a unique distinct M serotype.
研究了利用rRNA基因限制性片段长度多态性分析（核糖体分型）对化脓性链球菌菌株进行亚型分类。对68株化脓性链球菌菌株进行了特征分析，其中包括17株参考菌株以及51株分别从血液、急性或慢性咽炎及食源性暴发中分离得到的菌株，通过测定它们的rDNA限制性片段长度多态性图谱和血清型（T和M）来进行分析。用5种选定的限制性内切酶切割总DNA，然后用一段与16S rRNA基因杂交的1063 bp地高辛标记片段进行探针杂交。SacI和XhoI分别产生了15种和9种不同的图谱，另外3种限制性内切酶每种都产生了5种图谱。使用SacI-XhoI组合，在68株分离株中总共获得了21种不同的核糖体分型。其他限制性内切酶的结果并未增加这个数量。所有测试菌株均可分型。每次食源性暴发中的所有分离株都属于同一核糖体分型，每个患有慢性咽炎的儿童的所有分离株（治疗前和治疗后）也都属于同一核糖体分型，提示存在抗生素治疗失败的情况。使用Hunter-Gaston公式为47株在流行病学上无关联的分离株计算了鉴别指数。当使用SacI-XhoI组合时，该指数达到0.955，表明该分型方法具有良好的鉴别能力。因此，核糖体分型被证明是一种对化脓性链球菌进行亚型分类的有意义的分子方法。此外，核糖体分型和血清分型之间存在一定相关性，因为几种核糖体分型与独特的不同M血清型相关。
