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[志贺氏菌中CRISPR的检测及其与耐药性的关系]

[Detection of CRISPR and its relationship to drug resistance in Shigella].

作者信息

Wang Linlin, Wang Yingfang, Duan Guangcai, Xue Zerun, Guo Xiangjiao, Wang Pengfei, Xi Yuanlin, Yang Haiyan

出版信息

Wei Sheng Wu Xue Bao. 2015 Apr 4;55(4):476-83.

Abstract

OBJECTIVE

To detect clustered regularly interspaced short palindromic repeats (CRISPR) in Shigella, and to analyze its relationship to drug resistance.

METHODS

Four pairs of primers were used for the detection of convincing CRISPR structures CRISPR-S2 and CRISPR-S4, questionable CRISPR structures CRISPR-S1 and CRISPR-S3 in 60 Shigella strains. All primers were designed using sequences in CRISPR database. CRISPR Finder was used to analyze CRISPR and susceptibilities of Shigella strains were tested by agar diffusion method. Furthermore, we analyzed the relationship between drug resistance and CRISPR-S4.

RESULTS

The positive rate of convincing CRISPR structures was 95%. The four CRISPR loci formed 12 spectral patterns (A-L), all of which contained convincing CRISPR structures except type K. We found one new repeat and 12 new spacers. The multi-drug resistance rate was 53. 33% . We found no significant difference between CRISPR-S4 and drug resistant. However, the repeat sequence of CRISPR-S4 in multi- or TE-resistance strains was mainly R4.1 with AC deletions in the 3' end, and the spacer sequences of CRISPR-S4 in multi-drug resistance strains were mainly Sp5.1, Sp6.1 and Sp7.

CONCLUSION

CRISPR was common in Shigella. Variations df repeat sequences and diversities of spacer sequences might be related to drug resistance in Shigella.

摘要

目的

检测志贺菌中规律成簇间隔短回文重复序列(CRISPR),并分析其与耐药性的关系。

方法

使用4对引物检测60株志贺菌中可信的CRISPR结构CRISPR-S2和CRISPR-S4、可疑的CRISPR结构CRISPR-S1和CRISPR-S3。所有引物均根据CRISPR数据库中的序列设计。使用CRISPR Finder分析CRISPR,并采用琼脂扩散法检测志贺菌菌株的药敏情况。此外,分析耐药性与CRISPR-S4之间的关系。

结果

可信CRISPR结构的阳性率为95%。4个CRISPR位点形成12种谱型(A-L),除K型外均包含可信的CRISPR结构。发现1个新的重复序列和12个新的间隔序列。多重耐药率为53.33%。未发现CRISPR-S4与耐药性之间存在显著差异。然而,多重耐药或耐替加环素菌株中CRISPR-S4的重复序列主要为R4.1,3'端有AC缺失,多重耐药菌株中CRISPR-S4的间隔序列主要为Sp5.1、Sp6.1和Sp7。

结论

CRISPR在志贺菌中普遍存在。重复序列的变异和间隔序列的多样性可能与志贺菌的耐药性有关。

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Detection and analysis of CRISPRs of Shigella.志贺氏菌CRISPRs的检测与分析
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