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感染多重耐药临床大肠杆菌菌株的新型裂解性噬菌体的分离与鉴定

Isolation and characterization of novel lytic bacteriophages that infect multi drug resistant clinical strains of Escherichia coli.

作者信息

Padmesh Sudhaker, Singh Aditi, Chopra Sidharth, Sen Manodeep, Habib Saman, Shrivastava Deepti, Johri Parul

机构信息

Amity Institute of Biotechnology, Amity University Uttar Pradesh, Lucknow Campus, Gomti Nagar Extension, Lucknow, 226028, India.

Division of Microbiology, CSIR-Central Drug Research Institute, Lucknow, 226031, India.

出版信息

Environ Sci Pollut Res Int. 2024 Oct;31(46):57327-57337. doi: 10.1007/s11356-023-28081-z. Epub 2023 Jun 22.

DOI:10.1007/s11356-023-28081-z
PMID:37347328
Abstract

The pathogenic strains of Escherichia coli (E. coli) are frequent cause of urinary tract infections including catheter-associated, soft tissue infections and sepsis. The growing antibiotic resistance in E. coli is a major health concern. Bacteriophages are specific for their bacterial host, thus providing a novel and effective alternatives. This study focuses on isolation of bacteriophages from urban sewage treatment plants. Initially 50 different bacteriophages have been isolated against non-resistant reference E. coli strain and fifty multidrug resistant clinical isolates of extraintestinal infections. Out of which only thirty-one lytic phages which gave clear plaques were further analysed for different physico-chemical aspects such as thermal inactivation, pH, effect of organic solvents and detergents. Two bacteriophages, ASEC2201 and ASEC2202, were selected for their ability to withstand temperature fluctuation from -20 to 62 °C and a pH range from 4 to 10. They also showed good survival (40-94%) in the presence of organic solvents like ethanol, acetone, DMSO and chloroform or ability to form plaques even after the treatment with detergents like SDS, CTAB and sarkosyl. Both efficiently killed reference strain and 40-44% of multidrug resistant clinical isolates of E. coli. Later ASEC2201 and ASEC2202 were subjected to morphological characterisation through transmission electron microscopy, which revealed them to be tailed phages. The genomic analysis confirmed them to be Escherichia phages which belonged to family Drexlerviridae of Caudovirales.

摘要

大肠杆菌(E. coli)的致病菌株是包括导管相关感染、软组织感染和败血症在内的尿路感染的常见病因。大肠杆菌中日益增长的抗生素耐药性是一个主要的健康问题。噬菌体对其细菌宿主具有特异性,因此提供了一种新颖且有效的替代方案。本研究的重点是从城市污水处理厂中分离噬菌体。最初,已针对非耐药参考大肠杆菌菌株和50株肠道外感染的多重耐药临床分离株分离出50种不同的噬菌体。其中,仅对产生清晰噬菌斑的31种裂解性噬菌体进行了进一步的不同物理化学方面的分析,如热灭活、pH值、有机溶剂和洗涤剂的影响。选择了两种噬菌体ASEC2201和ASEC2202,因为它们能够承受-20至62°C的温度波动以及4至10的pH范围。它们在乙醇、丙酮、二甲基亚砜和氯仿等有机溶剂存在下也表现出良好的存活率(40-94%),或者即使在用十二烷基硫酸钠、十六烷基三甲基溴化铵和 Sarkosyl 等洗涤剂处理后仍有形成噬菌斑的能力。两者均能有效杀死参考菌株以及40-44%的大肠杆菌多重耐药临床分离株。后来,通过透射电子显微镜对ASEC2201和ASEC2202进行了形态学表征,结果显示它们是有尾噬菌体。基因组分析证实它们是属于有尾噬菌体目德雷克斯勒病毒科的大肠杆菌噬菌体。

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