生物技术通报 ›› 2022, Vol. 38 ›› Issue (9): 127-135.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1549

• 细菌耐药性专题(专题主编: 刘雅红 教授 孙坚 教授) • 上一篇    下一篇

一株新的多重耐药福氏志贺菌噬菌体生物学特性及基因组分析

文畅(), 刘晨, 卢诗韵, 许忠兵, 艾超凡, 廖汉鹏(), 周顺桂   

  1. 福建省土壤环境健康与调控重点实验室 福建农林大学资源与环境学院,福州 350002
  • 收稿日期:2021-12-14 出版日期:2022-09-26 发布日期:2022-10-11
  • 作者简介:文畅,女,硕士研究生,研究方向:环境病毒与抗生素耐药性;E-mail: 2742707020@qq.com
  • 基金资助:
    国家自然科学基金项目(31972521);岭南现代农业实验室科研项目(NT2021010)

Biological Characteristics and Genome Analysis of a Novel Multidrug-resistant Shigella flexneri Phage

WEN Chang(), LIU Chen, LU Shi-yun, XU Zhong-bing, AI Chao-fan, LIAO Han-peng(), ZHOU Shun-gui   

  1. Fujian Provincial Key Laboratory of Soil Environmental Health and Regulation,College of Resources and Environment,Fujian Agriculture and Forestry University,Fuzhou 350002
  • Received:2021-12-14 Published:2022-09-26 Online:2022-10-11

摘要:

抗生素的大量使用和滥用造成细菌耐药性问题愈发严峻,噬菌体疗法作为一种精准治疗多重耐药病原菌的有效方法开始被人们日益重视。采用双层琼脂平板法从活性污泥中分离出多重耐药福氏志贺菌(Shigella flexneri)噬菌体P003,鉴定为肌尾噬菌体科(Myoviridae)。通过生物学特性测定表明P003的最佳感染复数为10,潜伏期约10 min;在4-70℃、pH 3.0-10.0的条件下保持活性。全基因组测序表明,噬菌体基因组长约68 721 bp,GC含量46.14%,预测编码99个开放阅读框(open reading frame,ORF),未发现已知耐药基因和毒力基因。全基因组多序列线性与进化树分析结果表明,P003与大肠杆菌噬菌体有较近的亲缘关系,但不能侵染大肠杆菌。从活性污泥原位分离到一株新的多重耐药福氏志贺菌噬菌体P003,为利用噬菌体疗法防治多重耐药病原菌S. flexneri感染提供了新的菌种资源。

关键词: 噬菌体, 福氏志贺菌, 抗生素耐药性, 生物学特性, 全基因组分析

Abstract:

Antibiotic resistance of bacteria caused by over use and abuse of antibiotics has been more and more seriously,and phage therapy as an precision treatment to treat multi-drug-resistant pathogens has being received more attentions. Double-layer agar plate method was applied to isolate a phage P300,a multi-drug resistant Shigella flexneri,from activated sludge and identified as Myoviridae. The measurement of biological characteristics demonstrated that the optimal multiplicity of infection(MOI)of phage P003 was 10,and the latent period was 10 min. Phage P003 remained active under the conditions of 4-70℃ and pH 3.0-10.0. The whole genome sequencing of phage P003 revealed that the length was 68 721 bp,contained 99 ORFs and 46.14% GC content,and no known drug resistance genes and virulence genes were discovered. Whole genome multi-sequence linear comparison and phylogenetic tree analysis showed that P003 was highly similar to Escherichia coli phage,but it cannot infect E. coli,indicating that P003 was a novel phage. In sum,a novel multidrug resistant phage S. flexneri P003 is isolated from active sludge,which may provide a new strain resource for the treatment of multidrug-resistant pathogen S. flexneri infection by phage therapy in the future.

Key words: phage, Shigella flexneri, antibiotic resistance, biological characteristics, whole genome sequencing