Manos Jim, Arthur Jonathan, Rose Barbara, Tingpej Pholawat, Fung Carina, Curtis Michelle, Webb Jeremy S, Hu Honghua, Kjelleberg Staffan, Gorrell Mark D, Bye Peter, Harbour Colin
Department of Infectious Diseases and Immunology, University of Sydney, Sydney, Australia.
Sydney Bioinformatics, University of Sydney, Sydney, Australia.
J Med Microbiol. 2008 Dec;57(Pt 12):1454-1465. doi: 10.1099/jmm.0.2008/005009-0.
Transmissible Pseudomonas aeruginosa clones potentially pose a serious threat to cystic fibrosis (CF) patients. The AES-1 clone has been found to infect up to 40 % of patients in five CF centres in eastern Australia. Studies were carried out on clonal and non-clonal (NC) isolates from chronically infected CF patients, and the reference strain PAO1, to gain insight into the properties of AES-1. The transcriptomes of AES-1 and NC isolates, and of PAO1, grown planktonically and as a 72 h biofilm were compared using PAO1 microarrays. Microarray data were validated using real-time PCR. Overall, most differentially expressed genes were downregulated. AES-1 differentially expressed bacteriophage genes, novel motility genes, and virulence and quorum-sensing-related genes, compared with both PAO1 and NC. AES-1 but not NC biofilms significantly downregulated aerobic respiration genes compared with planktonic growth, suggesting enhanced anaerobic/microaerophilic growth by AES-1. Biofilm measurement showed that AES-1 formed significantly larger and thicker biofilms than NC or PAO1 isolates. This may be related to expression of the gene PA0729, encoding a biofilm-enhancing bacteriophage, identified by PCR in all AES-1 but few NC isolates (n=42). Links with the Liverpool epidemic strain included the presence of PA0729 and the absence of the bacteriophage gene cluster PA0632-PA0639. No common markers were found with the Manchester strain. No particular differentially expressed gene in AES-1 could definitively be ascribed a role in its infectivity, thus increasing the likelihood that AES-1 infectivity is multi-factorial and possibly involves novel genes. This study extends our understanding of the transcriptomic and genetic differences between clonal and NC strains of P. aeruginosa from CF lung.
可传播的铜绿假单胞菌克隆可能对囊性纤维化(CF)患者构成严重威胁。已发现AES-1克隆在澳大利亚东部的五个CF中心感染了多达40%的患者。对来自慢性感染CF患者的克隆和非克隆(NC)分离株以及参考菌株PAO1进行了研究,以深入了解AES-1的特性。使用PAO1微阵列比较了AES-1和NC分离株以及PAO1在浮游生长和形成72小时生物膜时的转录组。微阵列数据通过实时PCR进行验证。总体而言,大多数差异表达基因被下调。与PAO1和NC相比,AES-1差异表达噬菌体基因、新型运动基因以及毒力和群体感应相关基因。与浮游生长相比,AES-1生物膜而非NC生物膜显著下调需氧呼吸基因,这表明AES-1增强了厌氧/微需氧生长。生物膜测量显示,AES-1形成的生物膜比NC或PAO1分离株显著更大更厚。这可能与基因PA0729的表达有关,该基因编码一种增强生物膜的噬菌体,通过PCR在所有AES-1分离株中均有发现,但在少数NC分离株(n = 42)中未发现。与利物浦流行菌株的联系包括PA0729的存在以及噬菌体基因簇PA0632 - PA0639的缺失。未发现与曼彻斯特菌株的共同标记。AES-1中没有特定的差异表达基因可以明确归因于其感染性,因此增加了AES-1感染性是多因素的且可能涉及新基因的可能性。这项研究扩展了我们对CF肺部铜绿假单胞菌克隆和NC菌株之间转录组和遗传差异的理解。