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应用抑制性消减杂交技术鉴定和分析临床 1A 型肠炎耶尔森菌的毒力基因。

Identification and distribution of putative virulence genes in clinical strains of Yersinia enterocolitica biovar 1A by suppression subtractive hybridization.

机构信息

Microbial Pathogenicity Laboratory, Department of Microbiology, University of Delhi South Campus, New Delhi, India.

出版信息

J Appl Microbiol. 2012 Nov;113(5):1263-72. doi: 10.1111/j.1365-2672.2012.05427.x. Epub 2012 Sep 5.

Abstract

AIMS

To detect putative virulence genes in clinical strains of Yersinia enterocolitica biovar 1A by suppression subtractive hybridization between two closely related strains of clinical and nonclinical origin having the same serotype (O:6,30-6,31).

METHODS AND RESULTS

Suppression Subtractive Hybridization (SSH) was used to identify genomic differences between clinical (serotype O:6,30-6,31, from diarrhoeic human stools) and nonclinical (serotype O:6,30-6,31, from wastewater) strains of Y. enterocolitica biovar 1A. Following genomic subtraction and DNA sequencing, nine DNA sequences that were present only in clinical biovar 1A strains were identified. The sequences identified using SSH showed similarity to conserved hypothetical proteins, proteins related to iron acquisition and haemin storage, type 1 secretion proteins, flagellar hook proteins, exported protein and ABC transport system. All these sequences showed high similarity with Y. enterocolitica 8081 (biovar 1B). The distribution of these genes was further analysed using PCR in 26 clinical strains of Y. enterocolitica biovar 1A. The results revealed that the distribution of these genes was not uniform.

CONCLUSIONS

Genes related to iron acquisition and storage, and flagellar proteins might be responsible for virulence of some of the clinical strains of Y. enterocolitica biovar 1A.

SIGNIFICANCE AND IMPACT OF THE STUDY

Genes identified in this study might be useful in understanding the pathogenic potential of clinical strains of Y. enterocolitica biovar 1A.

摘要

目的

通过对具有相同血清型(O:6,30-6,31)的临床和非临床来源的两种密切相关菌株之间的抑制性消减杂交,检测肠侵袭性大肠杆菌 1A 生物型临床株中的假定毒力基因。

方法和结果

采用抑制性消减杂交(SSH)技术来鉴定肠侵袭性大肠杆菌 1A 生物型临床株(来自腹泻人类粪便,血清型 O:6,30-6,31)和非临床株(来自废水,血清型 O:6,30-6,31)之间的基因组差异。经过基因组消减和 DNA 测序,鉴定出仅存在于临床 1A 生物型菌株中的 9 个 DNA 序列。使用 SSH 鉴定的序列与保守的假设蛋白、与铁摄取和血红素储存相关的蛋白、1 型分泌蛋白、鞭毛钩蛋白、分泌蛋白和 ABC 转运系统具有相似性。所有这些序列与肠侵袭性大肠杆菌 8081(1B 生物型)具有高度相似性。进一步使用 PCR 在 26 株临床分离的肠侵袭性大肠杆菌 1A 生物型菌株中分析这些基因的分布。结果表明,这些基因的分布并不均匀。

结论

与铁摄取和储存以及鞭毛蛋白相关的基因可能是某些临床分离的肠侵袭性大肠杆菌 1A 生物型菌株毒力的原因。

本研究鉴定的基因可能有助于理解临床分离的肠侵袭性大肠杆菌 1A 生物型菌株的致病潜力。

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