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应用聚类规则间隔短回文重复序列分析解读孕妇阴道带菌的 B 族链球菌

Group B Streptococcus Vaginal Carriage in Pregnant Women as Deciphered by Clustered Regularly Interspaced Short Palindromic Repeat Analysis.

机构信息

Université François Rabelais de Tours, UMR1282 Infectiologie et Santé Publique, Tours, France.

INRA, UMR1282 Infectiologie et Santé Publique, Nouzilly, France.

出版信息

J Clin Microbiol. 2018 May 25;56(6). doi: 10.1128/JCM.01949-17. Print 2018 Jun.

Abstract

We evaluated the diversity of group B (GBS) vaginal carriage populations in pregnant women. For this purpose, we studied each isolate present in a primary culture of a vaginal swab using a new approach based on clustered regularly interspaced short palindromic repeats (CRISPR) locus analysis. To evaluate the CRISPR array composition rapidly, a restriction fragment length polymorphism (RFLP) analysis was performed. For each different pattern observed, the CRISPR array was sequenced and capsular typing and multilocus sequence typing (MLST) were performed. A total of 970 isolates from 10 women were analyzed by CRISPR-RFLP. Each woman carrying GBS isolates presented one to five specific "personal" patterns. Five women showed similar isolates with specific and unique restriction patterns, suggesting the carriage of a single GBS clone. Different patterns were observed among isolates from the other five women. For three of these, CRISPR locus sequencing highlighted low levels of internal modifications in the locus backbone, whereas there were high levels of modifications for the last two women, suggesting the carriage of two different clones. These two clones were closely related, having the same ancestral spacer(s), the same capsular type and, in one case, the same ST, but showed different antibiotic resistance patterns in pairs. Eight of 10 women were colonized by a single GBS clone, while two of them were colonized by two strains, leading to a risk of selection of more-virulent and/or more-resistant clones during antibiotic prophylaxis. This CRISPR analysis made it possible to separate isolates belonging to a single capsular type and sequence type, highlighting the greater discriminating power of this approach.

摘要

我们评估了孕妇群体 B 型(GBS)阴道携带人群的多样性。为此,我们使用一种新的基于簇状规律间隔短回文重复序列(CRISPR)基因座分析的方法,对阴道拭子的初次培养中存在的每个分离株进行研究。为了快速评估 CRISPR 基因座组成,我们进行了限制性片段长度多态性(RFLP)分析。对于观察到的每种不同模式,对 CRISPR 基因座进行测序,并进行荚膜分型和多位点序列分型(MLST)。通过 CRISPR-RFLP 分析了来自 10 名女性的 970 个分离株。携带 GBS 分离株的每位女性均呈现出一到五个特定的“个人”模式。五名女性携带具有特定且独特的限制模式的相似分离株,表明携带单一 GBS 克隆。来自其他五名女性的分离株观察到不同的模式。对于其中三个,CRISPR 基因座测序突出了基因座骨干内部修饰的低水平,而对于最后两名女性,修饰水平较高,表明携带两个不同的克隆。这两个克隆密切相关,具有相同的祖先间隔子,相同的荚膜型,在一种情况下,相同的 ST,但在配对时显示出不同的抗生素耐药模式。10 名女性中有 8 名被单个 GBS 克隆定植,而其中 2 名被两个菌株定植,这导致在抗生素预防期间选择更具毒力和/或更具耐药性的克隆的风险增加。这项 CRISPR 分析使我们能够分离属于单个荚膜型和序列型的分离株,突出了这种方法具有更高的区分能力。

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