论著·基础研究

耐碳青霉烯类鲍曼不动杆菌噬菌体SH-Ab15599的生物学特性及基因组学分析

  • 季佳炜 1 ,
  • 2 ,
  • 王睿 1 ,
  • 罗婷婷 4 ,
  • 徐梦莎 1 ,
  • 郭晓奎 1 ,
  • 胡付品 3 ,
  • 李敏 2 ,
  • 何平 1
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  • 1. 上海交通大学基础医学院免疫学与微生物学系,上海 200025;2. 上海交通大学医学院附属仁济医院检验科,上海 200127;3.复旦大学附属华山医院抗生素研究所,上海 200040;4. 浙江医药高等专科学校制药工程学院,宁波 315100
季佳炜(1988—),男,硕士生;电子信箱: randll-jjw@163.com。

网络出版日期: 2018-12-15

基金资助

国家自然科学基金(81471908)

Characterization and genome sequencing of SH-Ab 15599, a novel bacteriophage targeting carbapenem-resistant Acinetobacter baumannii

  • JI Jia-wei1 ,
  • 2 ,
  • WANG Rui1 ,
  • LUO Ting-ting4 ,
  • XU Meng-sha1 ,
  • GUO Xiao-kui1 ,
  • HU Fu-pin3 ,
  • LI Min2 ,
  • HE Ping1
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  • 1. Department of Immunology and Microbiology, Shanghai Jiao Tong University College of Basic Medical Sciences, Shanghai 200025, China; 2. Department of Laboratory Medicine, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China; 3. Institute of Antibiotics, Huashan Hospital, Fudan University, Shanghai 200040, China; 4. Department of Pharmaceutical Engineering, Zhejiang Pharmaceutical College, Ningbo 315100, China

Online published: 2018-12-15

Supported by

National Natural Science Foundation of China, 81471908

摘要

目的 ·寻找具有感染耐碳青霉烯类鲍曼不动杆菌 (carbapenem-resistant Acinetobacter baumannii, CRAB)潜力的烈性噬菌体,进行生物学特性及基因组学分析。方法 ·采集污水,富集分离纯化得到噬菌体,通过电子显微镜观察噬菌体形态,研究其理化特征。抽取噬菌体 DNA,进行测序和基因分析。结果 ·分离得到 l 株鲍曼不动杆菌噬菌体,命名为 SH-Ab 15599。电子显微镜观察结果显示该噬菌体为肌尾噬菌体。 SH-Ab 15599能够裂解 51株 CRAB临床分离株中的 15株 (29.4%);在 4~ 50℃和 pH值 6~ 9的环境下, 1 h内能保持活性;与宿主菌共培养 5 min后,吸附率约为 90%;其潜伏期约 10 min,裂解期约 20 min,每个细胞裂解量约 43 pfu。在体外实验中,宿主菌的生长受到抑制。 SH-Ab 15599的基因组为双链线状 DNA,含 143 204 bp,GC含量为 38.45%。结论 · SH-Ab 15599是一株全新的噬菌体,具有治疗 CRAB感染的潜力。

本文引用格式

季佳炜 1 , 2 , 王睿 1 , 罗婷婷 4 , 徐梦莎 1 , 郭晓奎 1 , 胡付品 3 , 李敏 2 , 何平 1 . 耐碳青霉烯类鲍曼不动杆菌噬菌体SH-Ab15599的生物学特性及基因组学分析[J]. 上海交通大学学报(医学版), 2018 , 38(11) : 1283 . DOI: 10.3969/j.issn.1674-8115.2018.11.002

Abstract

Objective · To find lytic bacteriophages which have potential to combat carbapenem-resistant Acinetobacter baumannii (CRAB) infections and analyze the biological characterizations and genomic sequences of the phages. Methods · The phages were isolated sewage followedamplification and purification. They were characterizedthe transmission electron microscopy, the host specificity, the biological characterizations, the lysis assay in vitro and the genome sequence analysis. Results · A novel bacteriophage, SH-Ab 15599, was isolated sewage. The electron microscope revealed SH-Ab 15599 belongs to Myoviridae. Among the 51 clinical CRAB isolates tested, 15 (29.4%) were lysedthe phage. SH-Ab 15599 was relatively sat temperature range 4 ℃ to 50 ℃ or between pH 6 and 9. About 90% of the phage was approximately adsorbed to the host CRAB isolate in 5 min. The one-step growth curve showed that the latent period was about 10 min and the growth period was about 20 min. The burst size was estimated at 43 pfu per infected cell. The growth of the host CRAB isolate was inhibitedthe phage in vitro. The genome of SH-Ab 15599 was a linear, 143 204 bp double-stranded DNA with GC content of 38.45%. Conclusion · SH-Ab 15599 is a novel phage targeting CRAB and has potential as an alternative against CRAB infections in the future.
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