Beauruelle Clémence, Treluyer Ludovic, Pastuszka Adeline, Cochard Thierry, Lier Clément, Mereghetti Laurent, Glaser Philippe, Poyart Claire, Lanotte Philippe
Département de Bactériologie-Virologie, Hygiène Hospitalière et Parasitologie-Mycologie, Centre Hospitalier Régional Universitaire (CHRU) de Brest, Brest, France.
Inserm, EFS, UMR 1078, GGB, Universitè de Bretagne Occidentale, Brest, France.
Front Microbiol. 2021 Jul 19;12:675597. doi: 10.3389/fmicb.2021.675597. eCollection 2021.
We explored the relevance of a Clustered regularly interspaced short palindromic repeats (CRISPR)-based genotyping tool for typing and we compared this method to current molecular methods [multi locus sequence typing (MLST) and capsular typing]. To this effect, we developed two CRISPR marker schemes (using 94 or 25 markers, respectively). Among the 255 isolates tested, 229 CRISPR profiles were obtained. The 94 and 25 markers made it possible to efficiently separate isolates with a high diversity index (0.9947 and 0.9267, respectively), highlighting a high discriminatory power, superior to that of both capsular typing and MLST (diversity index of 0.9017 for MLST). This method has the advantage of being correlated with MLST [through analysis of the terminal direct repeat (TDR) and ancestral spacers] and to possess a high discriminatory power (through analysis of the leader-end spacers recently acquired, which are the witnesses of genetic mobile elements encountered by the bacteria). Furthermore, this "one-shot" approach presents the benefit of much-reduced time and cost in comparison with MLST. On the basis of these data, we propose that this method could become a reference method for group B (GBS) typing.
我们探究了一种基于成簇规律间隔短回文重复序列(CRISPR)的基因分型工具在分型方面的相关性,并将该方法与当前的分子方法[多位点序列分型(MLST)和荚膜分型]进行了比较。为此,我们开发了两种CRISPR标记方案(分别使用94个或25个标记)。在测试的255株分离株中,获得了229种CRISPR图谱。94个和25个标记能够有效地分离具有高多样性指数的分离株(分别为0.9947和0.9267),突出显示了其高鉴别力,优于荚膜分型和MLST(MLST的多样性指数为0.9017)。该方法的优点是与MLST相关[通过对末端直接重复序列(TDR)和祖先间隔序列的分析],并且具有高鉴别力(通过对最近获得的前导端间隔序列的分析,这些间隔序列是细菌遇到的遗传移动元件的见证)。此外,与MLST相比,这种“一次性”方法具有大大减少时间和成本的优势。基于这些数据,我们建议该方法可成为B组链球菌(GBS)分型的参考方法。