Characterising the biology of novel lytic bacteriophages infecting multidrug resistant Klebsiella pneumoniae.
Characterising the biology of novel lytic bacteriophages infecting multidrug resistant Klebsiella pneumoniae.
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DOI：
10.1186/1743-422x-10-100
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发表时间：
2013-03-28
期刊：
影响因子：
4.8
通讯作者：
Kropinski AM
中科院分区：
文献类型：
--
作者：
Kęsik-Szeloch A;Drulis-Kawa Z;Weber-Dąbrowska B;Kassner J;Majkowska-Skrobek G;Augustyniak D;Lusiak-Szelachowska M;Zaczek M;Górski A;Kropinski AM
克雷伯氏菌属的成员是与医院感染相关的主要微生物病原体之一。这些物种中抗菌素耐药性的发生率增加，促使需要替代/联合治疗方案来帮助临床治疗。噬菌体疗法形成这些替代策略之一。利用电子显微镜、爆发大小、宿主范围、噬菌体颗粒对温度、氯仿、pH的敏感性以及噬菌体DNA的限制性消化来表征克雷伯氏菌。在32个分离的病毒中，8个属于肌尾病毒科，8个属于管尾病毒科，其余16个属于短尾病毒科。这些细菌的宿主范围针对254种临床肠杆菌科菌株进行了表征，包括产生超广谱β-内酰胺酶（ESBLs）的多重耐药克雷伯菌属分离株。根据其裂解潜力，进一步表征了其中六个噬菌体的爆发大小、物理化学性质和对限制性内切酶消化的敏感性。此外，5个基因已完全测序。鉴定了多种噬菌体编码的宿主抗性机制。Siphoviridae噬菌体基因组（KP 16和KP 36）含有少量的宿主限制性位点，类似于T7样噬菌体（KP 32）中发现的策略。此外，噬菌体KP 36编码其自身的DNA腺嘌呤甲基转移酶。φ KMV-样KP 34噬菌体对本研究中使用的所有核酸内切酶均敏感。检测到KP 34 DNA的Dam甲基化，尽管这是在不存在可识别的噬菌体编码的甲基转移酶的情况下。Myoviridae KP 15和KP 27都携带Dam和Dcm甲基转移酶基因以及先前研究中阐明的其他抗限制性机制。没有发现其他的抗限制机制，例如非典型核苷酸（hmC或葡糖基hmC），尽管肌尾病毒科噬菌体KP 27编码一种未知的抗限制机制，需要进一步研究。
Members of the genus Klebsiella are among the leading microbial pathogens associated with nosocomial infection. The increased incidence of antimicrobial resistance in these species has propelled the need for alternate/combination therapeutic regimens to aid clinical treatment. Bacteriophage therapy forms one of these alternate strategies. Electron microscopy, burst size, host range, sensitivity of phage particles to temperature, chloroform, pH, and restriction digestion of phage DNA were used to characterize Klebsiella phages. Of the 32 isolated phages eight belonged to the family Myoviridae, eight to the Siphoviridae whilst the remaining 16 belonged to the Podoviridae. The host range of these phages was characterised against 254 clinical Enterobacteriaceae strains including multidrug resistant Klebsiella isolates producing extended-spectrum beta-lactamases (ESBLs). Based on their lytic potential, six of the phages were further characterised for burst size, physicochemical properties and sensitivity to restriction endonuclease digestion. In addition, five were fully sequenced. Multiple phage-encoded host resistance mechanisms were identified. The Siphoviridae phage genomes (KP16 and KP36) contained low numbers of host restriction sites similar to the strategy found in T7-like phages (KP32). In addition, phage KP36 encoded its own DNA adenine methyltransferase. The φKMV-like KP34 phage was sensitive to all endonucleases used in this study. Dam methylation of KP34 DNA was detected although this was in the absence of an identifiable phage encoded methyltransferase. The Myoviridae phages KP15 and KP27 both carried Dam and Dcm methyltransferase genes and other anti-restriction mechanisms elucidated in previous studies. No other anti-restriction mechanisms were found, e.g. atypical nucleotides (hmC or glucosyl hmC), although Myoviridae phage KP27 encodes an unknown anti-restriction mechanism that needs further investigation.