Survey of Pseudomonas aeruginosa and its phages: de novo peptide sequencing as a novel tool to assess the diversity of worldwide collected viruses
Survey of Pseudomonas aeruginosa and its phages: de novo peptide sequencing as a novel tool to assess the diversity of worldwide collected viruses
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DOI：
10.1111/j.1462-2920.2008.01862.x
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发表时间：
2009-05-01
影响因子：
5.1
通讯作者：
Lavigne, Rob
中科院分区：
文献类型：
--
作者：
Ceyssens, Pieter-Jan;Noben, Jean-Paul;Lavigne, Rob
收集了15个新分离的感染条件致病菌铜绿假单胞菌的细菌，以研究它们的全球多样性和在噬菌体治疗中的潜力。这些粪便是在横跨四大洲的14个不同国家从自然环境和医院污水中取样的。它们都显示出独特的DNA和蛋白质谱，并在尾状病毒目的三个主要家族内形态上聚类为六个组。对122株AFLP基因型临床铜绿假单胞菌菌株（其中49株是在比利时鲁汶大学医院新分离的）的文库进行的广泛宿主范围研究表明，87%的菌株被裂解。外膜突变体的感染分析确定了10个IV型菌毛依赖性。通过使用串联基质辅助激光解吸/电离飞行时间技术对主要病毒体蛋白质进行从头肽测序，收集了关于病毒进化相关性的更详细信息。首次将该技术应用于病毒，在不需要基因组内容和/或电子显微镜成像的先验知识的情况下鉴定了7组密切相关的病毒。本研究证明了铜绿假单胞菌的流行性种群结构和铜绿假单胞菌噬菌体物种的全球传播，并指出两种临床上占优势的、广泛分布的铜绿假单胞菌菌株对噬菌体攻击的抗性。
A collection of 15 newly isolated (bacterio)phages infecting the opportunistic pathogen Pseudomonas aeruginosa was established to investigate their global diversity and potential in phage therapy. These phages were sampled in 14 different countries traversing four continents, from both natural environments and hospital sewage. They all display unique DNA and protein profiles and cluster morphologically into six groups within the three major families of the Caudovirales. Extensive host range studies on a library of 122 AFLP-genotyped clinical P. aeruginosa strains (of which 49 were newly isolated at the University Hospital of Leuven, Belgium) showed that the phages lysed 87% of the strains. Infection analysis of outer membrane mutants identified 10 phages as type IV pili-dependent. More detailed information about the evolutionary relatedness of the phages was gathered by de novo peptide sequencing of major virion proteins using tandem Matrix-Assisted Laser Desorption/Ionization Time of Flight technology. Applying this technique for the first time to viruses, seven groups of closely related phages were identified without the need of prior knowledge of genome content and/or electron microscopic imaging. This study demonstrates both the epidemic population structure of P. aeruginosa and the global spread of P. aeruginosa phage species, and points at the resistance of two clinically predominant, widespread P. aeruginosa strains against phage attack.