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新型嗜麦芽窄食单胞菌巨型噬菌体phiSMA5的分离与鉴定

Isolation and characterization of novel giant Stenotrophomonas maltophilia phage phiSMA5.

作者信息

Chang Hsiao-Chuan, Chen Chiy-Rong, Lin Juey-Wen, Shen Gwan-Han, Chang Kai-Ming, Tseng Yi-Hsiung, Weng Shu-Fen

机构信息

Institute of Molecular Biology, National Chung Hsing University, Taichung 402, Taiwan.

出版信息

Appl Environ Microbiol. 2005 Mar;71(3):1387-93. doi: 10.1128/AEM.71.3.1387-1393.2005.

Abstract

Stenotrophomonas maltophilia is one of the most prevalent opportunistic bacteria causing nosocomial infections. It has become problematic because most of the isolates are resistant to multiple antibiotics, and therefore, development of phage therapy has attracted strong attention. In this study, eight S. maltophilia phages were isolated from clinical samples including patient specimens, catheter-related devices, and wastewater. These phages can be divided into four distinct groups based on host range and digestibility of the phage DNAs with different restriction endonucleases. One of them, designated phiSMA5, was further characterized. Electron microscopy showed it resembled Myoviridae, with an isometric head (90 nm in diameter), a tail (90 nm long), a baseplate (25 nm wide), and short tail fibers. The phiSMA5 double-stranded DNA, refractory to digestion by most restriction enzymes, was tested and estimated to be 250 kb by pulsed-field gel electrophoresis. This genome size is second to that of the largest phage, phiKZ of Pseudomonas aeruginosa. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, 25 virion proteins were visualized. N-terminal sequencing of four of them suggested that each of them might have had its N terminus cleaved off. Among the 87 S. maltophilia strains collected in this study, only 61 were susceptible to phiSMA5, indicating that more phages are needed toward a phage therapy strategy. Since literature search yielded no information about S. maltophilia phages, phiSMA5 appears to be the first reported.

摘要

嗜麦芽窄食单胞菌是引起医院感染的最常见的机会致病菌之一。由于大多数分离株对多种抗生素耐药,它已成为一个难题,因此,噬菌体疗法的开发引起了广泛关注。在本研究中,从包括患者标本、导管相关装置和废水在内的临床样本中分离出8株嗜麦芽窄食单胞菌噬菌体。根据宿主范围和噬菌体DNA对不同限制性内切酶的消化率,这些噬菌体可分为4个不同的组。其中一个命名为phiSMA5的噬菌体被进一步鉴定。电子显微镜显示它类似于肌病毒科,有一个等轴状头部(直径90nm)、一条尾部(90nm长)、一个基板(25nm宽)和短尾丝。phiSMA5双链DNA对大多数限制性酶具有抗性,经脉冲场凝胶电泳检测估计为250kb。这个基因组大小仅次于最大的噬菌体——铜绿假单胞菌的phiKZ。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中,可见25种病毒体蛋白。对其中4种蛋白进行N端测序表明,它们的N端可能都被切除了。在本研究收集的87株嗜麦芽窄食单胞菌菌株中,只有61株对phiSMA5敏感,这表明噬菌体疗法策略需要更多的噬菌体。由于文献检索未获得关于嗜麦芽窄食单胞菌噬菌体的信息,phiSMA5似乎是首次报道。

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