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通过单核苷酸多态性和脉冲场凝胶电泳基因分型分析铜绿假单胞菌群体的结构

Structure of Pseudomonas aeruginosa populations analyzed by single nucleotide polymorphism and pulsed-field gel electrophoresis genotyping.

作者信息

Morales Gracia, Wiehlmann Lutz, Gudowius Peter, van Delden Christian, Tümmler Burkhard, Martínez José Luis, Rojo Fernando

机构信息

Departamento de Biotecnología Microbiana, Centro Nacional de Biotecnología, CSIC, Campus de la UAM, Cantoblanco, 28049 Madrid, Spain.

出版信息

J Bacteriol. 2004 Jul;186(13):4228-37. doi: 10.1128/JB.186.13.4228-4237.2004.

Abstract

Pseudomonas aeruginosa has a wide ecological distribution that includes natural habitats and clinical settings. To analyze the population structure and distribution of P. aeruginosa, a collection of 111 isolates of diverse habitats and geographical origin, most of which contained a genome with a different SpeI macrorestriction profile, was typed by restriction fragment length polymorphism based on 14 single nucleotide polymorphisms (SNPs) located at seven conserved loci of the core genome (oriC, oprL, fliC, alkB2, citS, oprI, and ampC). The combination of these SNPs plus the type of fliC present (a or b) allowed the assignment of a genetic fingerprint to each strain, thus providing a simple tool for the discrimination of P. aeruginosa strains. Thirteen of the 91 identified SNP genotypes were found in two or more strains. In several cases, strains sharing their SNP genotype had different SpeI macrorestriction profiles. The highly virulent CHA strain shared its SNP genotype with other strains that had different SpeI genotypes and which had been isolated from nonclinical habitats. The reference strain PAO1 also shared its SNP genotype with other strains that had different SpeI genotypes. The P. aeruginosa chromosome contains a conserved core genome and variable amounts of accessory DNA segments (genomic islands and islets) that can be horizontally transferred among strains. The fact that some SNP genotypes were overrepresented in the P. aeruginosa population studied and that several strains sharing an SNP genotype had different SpeI macrorestriction profiles supports the idea that changes occur at a higher rate in the accessory DNA segments than in the conserved core genome.

摘要

铜绿假单胞菌具有广泛的生态分布,包括自然栖息地和临床环境。为了分析铜绿假单胞菌的种群结构和分布,对111株来自不同栖息地和地理来源的菌株进行了研究,其中大多数菌株的基因组具有不同的SpeI酶切图谱,基于位于核心基因组7个保守位点(oriC、oprL、fliC、alkB2、citS、oprI和ampC)的14个单核苷酸多态性(SNP),通过限制性片段长度多态性进行分型。这些SNP的组合加上存在的fliC类型(a或b),可为每个菌株赋予一个遗传指纹,从而提供了一种简单的工具来区分铜绿假单胞菌菌株。在91个已鉴定的SNP基因型中,有13个在两个或更多菌株中被发现。在几种情况下,共享SNP基因型的菌株具有不同的SpeI酶切图谱。高毒力的CHA菌株与其他具有不同SpeI基因型且从非临床栖息地分离出的菌株共享其SNP基因型。参考菌株PAO1也与其他具有不同SpeI基因型的菌株共享其SNP基因型。铜绿假单胞菌染色体包含一个保守的核心基因组和可变数量的辅助DNA片段(基因组岛和小岛),这些片段可在菌株间水平转移。在研究的铜绿假单胞菌种群中,某些SNP基因型出现频率过高,且一些共享SNP基因型的菌株具有不同的SpeI酶切图谱,这一事实支持了辅助DNA片段中的变化比保守核心基因组中的变化发生频率更高的观点。

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