Koessler Thibaud, Francois Patrice, Charbonnier Yvan, Huyghe Antoine, Bento Manuela, Dharan Sasi, Renzi Gesuele, Lew Daniel, Harbarth Stephan, Pittet Didier, Schrenzel Jacques
Genomic Research Laboratory, Service of Infectious Diseases, University of Geneva Hospitals, CH-1211 Geneva 14, Switzerland.
J Clin Microbiol. 2006 Mar;44(3):1040-8. doi: 10.1128/JCM.44.3.1040-1048.2006.
Until recently, methicillin-resistant Staphylococcus aureus (MRSA) was considered the prototype of a hospital-acquired bacterial pathogen. However, recent reports have shown that MRSA has now emerged in the community. Characterization of specific markers for distinguishing the origin of isolates could contribute to improved knowledge of MRSA epidemiology. The release of whole-genome sequences of hospital- and community-acquired S. aureus strains allowed the development of whole-genome content analysis techniques, including microarrays. We developed a microarray composed of 8,191 open reading frame-specific oligonucleotides covering >99% of the four sequenced S. aureus genomes (N315, Mu50, MW2, and COL) to evaluate gene contents of hospital- and community-onset S. aureus strains. In parallel, pulsed-field gel electrophoresis, variable number of tandem repeats, antibiogram, staphylococcal cassette chromosome-mec element typing, and presence of the Panton-Valentine leukocidin gene were evaluated in a collection of 15 clinical isolates. Clusters obtained with microarrays showed a high degree of similarity with those obtained by pulsed-field gel electrophoresis or variable number of tandem repeats. Clusters clearly segregated hospital-onset strains from community-onset strains. Moreover, the microarray approach allowed definition of novel marker genes and chromosomal regions specific for given groups of isolates, thus providing better discrimination and additional information compared to pulsed-field gel electrophoresis and variable number of tandem repeats. Finally, the comparative genome hybridization approach unraveled the occurrence of multiple horizontal transfer events leading to community-onset MRSA as well as the need for a specific genetic background in recipient strains for both the acquisition and the stability of the mec element.
直到最近,耐甲氧西林金黄色葡萄球菌(MRSA)还被认为是医院获得性细菌病原体的典型代表。然而,最近的报告显示,MRSA现已在社区中出现。鉴定区分分离株来源的特定标志物有助于增进对MRSA流行病学的了解。医院获得性和社区获得性金黄色葡萄球菌菌株全基因组序列的公布使得包括微阵列在内的全基因组内容分析技术得以发展。我们开发了一种由8191个开放阅读框特异性寡核苷酸组成的微阵列,覆盖了已测序的4个金黄色葡萄球菌基因组(N315、Mu50、MW2和COL)的>99%,以评估医院获得性和社区起病的金黄色葡萄球菌菌株的基因内容。同时,对15株临床分离株进行了脉冲场凝胶电泳、可变数目串联重复序列、抗菌谱、葡萄球菌盒式染色体mec元件分型以及杀白细胞素基因存在情况的评估。微阵列获得的聚类与脉冲场凝胶电泳或可变数目串联重复序列获得的聚类显示出高度相似性。聚类明显区分了医院起病菌株和社区起病菌株。此外,微阵列方法能够定义特定分离株群体特有的新标记基因和染色体区域,因此与脉冲场凝胶电泳和可变数目串联重复序列相比,能提供更好的区分度和更多信息。最后,比较基因组杂交方法揭示了导致社区起病MRSA的多次水平转移事件的发生,以及受体菌株中获得和稳定mec元件所需的特定遗传背景。