Phothichaisri Wichuda, Ounjai Puey, Phetruen Tanaporn, Janvilisri Tavan, Khunrae Pongsak, Singhakaew Sombat, Wangroongsarb Piyada, Chankhamhaengdecha Surang
Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok, Thailand.
Department of Biology, Faculty of Science, Mahidol University, Bangkok, Thailand.
Front Microbiol. 2018 Jul 31;9:1701. doi: 10.3389/fmicb.2018.01701. eCollection 2018.
is recognized as a problematic pathogen, causing severe enteric diseases including antibiotic-associated diarrhea and pseudomembranous colitis. The emergence of antibiotic resistant has driven a search for alternative anti-infection modalities. A promising strategy for controlling bacterial infection includes the use of bacteriophages and their gene products. Currently, knowledge of phages active against is still relatively limited by the fact that the isolation of phages for this organism is a technically demanding method since bacterial host themselves are difficult to culture. To isolate and characterize phages specific to , a genotoxic agent, mitomycin C, was used to induce temperate phages from 12 clinical isolates of . Five temperate phages consisting of ΦHR24, ΦHN10, ΦHN16-1, ΦHN16-2, and ΦHN50 were successfully induced and isolated. Spotting assays were performed against a panel of 92 isolates to screen for susceptible bacterial hosts. The results revealed that all the phages obtained in this work displayed a relatively narrow host range of 0-6.5% of the tested isolates. Electron microscopic characterization revealed that all isolated phages contained an icosahedral head connected to a long contractile tail, suggesting that they belonged to the family. Restriction enzyme analysis indicated that these phages possess unique double-stranded DNA genome. Further electron microscopic characterization revealed that the ΦHN10 absorbed to the bacterial surface via attachment to cell wall, potentially interacting with S-layer protein. Bacteriophages isolated from this study could lead to development of novel therapeutic agents and detection strategies for .
被认为是一种有问题的病原体,可引起严重的肠道疾病,包括抗生素相关性腹泻和假膜性结肠炎。抗生素耐药性的出现促使人们寻找替代的抗感染方式。控制细菌感染的一种有前景的策略包括使用噬菌体及其基因产物。目前,由于该生物体的噬菌体分离是一种技术要求较高的方法,因为细菌宿主本身难以培养,所以针对该菌的活性噬菌体的知识仍然相对有限。为了分离和鉴定特定于该菌的噬菌体,使用了一种基因毒性剂丝裂霉素C从该菌的12株临床分离株中诱导出温和噬菌体。成功诱导并分离出了由ΦHR24、ΦHN10、ΦHN16 - 1、ΦHN16 - 2和ΦHN50组成的五种温和噬菌体。针对一组92株该菌分离株进行了点滴试验,以筛选易感细菌宿主。结果显示,在这项工作中获得的所有该菌噬菌体的宿主范围相对较窄,仅为测试分离株的0 - 6.5%。电子显微镜表征显示,所有分离出的噬菌体都含有一个二十面体头部连接到一条长的收缩尾部,表明它们属于该菌噬菌体家族。限制性内切酶分析表明这些噬菌体拥有独特的双链DNA基因组。进一步的电子显微镜表征显示,ΦHN10通过附着在细胞壁上吸附到细菌表面,可能与S层蛋白相互作用。从本研究中分离出的噬菌体可能会导致针对该菌的新型治疗药物和检测策略的开发。