1株可裂解blaNDM-5及mcr-1阳性大肠杆菌噬菌体的分离与生物学特性

doi:10.11843/j.issn.0366-6964.2022.03.018

畜牧兽医学报 ›› 2022, Vol. 53 ›› Issue (3): 857-866.doi: 10.11843/j.issn.0366-6964.2022.03.018

1株可裂解bla_NDM-5及mcr-1阳性大肠杆菌噬菌体的分离与生物学特性

刘畅^1,2, 刘教^1,2, 陈进^1,2, 王勉之³, 熊文广^1,2, 曾振灵^1,2*

1. 华南农业大学兽医学院, 广州 510642;
2. 广东省兽药研制与安全评价重点实验室, 广州 510624;
3. 扬州大学兽医学院, 扬州 225009

收稿日期:2021-05-27 出版日期:2022-03-23 发布日期:2022-03-31
通讯作者: 曾振灵,主要从事兽医药理学与毒理学研究,E-mail:zlzeng@scau.edu.cn
作者简介:刘畅(1995-),女,湖北武汉人,硕士生,主要从事噬菌体及细菌耐药性研究,E-mail:2929224803@qq.com
基金资助:
国家自然科学基金项目(31872524);广东省普通高校省级重大科研项目(2017KZDXM006)

Isolation and Biological Characteristics of a Bacteriophage Capable of Lysing bla_NDM-5 and mcr-1 Positive Escherichia coli

LIU Chang^1,2, LIU Jiao^1,2, CHEN Jin^1,2, WANG Mianzhi³, XIONG Wenguang^1,2, ZENG Zhenling^1,2*

1. College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China;
2. Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety, Guangzhou 510642, China;
3. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China

Received:2021-05-27 Online:2022-03-23 Published:2022-03-31

摘要/Abstract

摘要： 旨在分离1株可裂解bla_NDM-5及mcr-1阳性大肠杆菌的噬菌体,探究该噬菌体的生物学特性,为防控bla_NDM-5及mcr-1阳性大肠杆菌感染提供新的技术思路。对分离出的噬菌体进行透射电镜观察、最佳感染复数、一步生长曲线等生物学特性试验及全基因组测序分析。结果显示:成功分离出1株大肠杆菌噬菌体vB_EcoM-ZQ1,该噬菌体属于有尾病毒目,肌尾病毒科,其最佳感染复数为0.1,潜伏期为20 min,裂解期为10 min,裂解量为92 PFU·cell^-1。在温度4~55℃、pH4~10条件下,该噬菌体都有较高的活性。其对多重耐药大肠杆菌有一定裂解作用,裂解率为25.5%。该噬菌体基因全长149 112 bp,GC含量为49.1%,共有184个ORF,其中70个ORF编码功能蛋白,有2个ORF编码tRNA,并未发现含有耐药基因与毒力因子;经BLAST比对发现,噬菌体与Shigella phage phiSboM-AG3有92.37%的相似性,基于噬菌体末端酶大亚基构建的同源进化树结果可得结论,噬菌体vB_EcoM-ZQ1和与沙门菌噬菌体pSal-SNUABM-02的亲缘关系最近。综上提示,噬菌体vB_EcoM-ZQ1,生物学特性稳定,基因结构完整,为后续bla_NDM-5及mcr-1阳性大肠杆菌感染的防控奠定了一定前期基础。

关键词: 噬菌体, 生物学特性, 大肠杆菌, bla_NDM-5, mcr-1

Abstract: This study aimed to isolate a phage that can lyse bla_NDM-5and mcr-1 positive Escherichia coli, explore the biological characteristics of the phage, and provide a certain theoretical basis for phage therapy. The isolated phage was observed by transmission electron microscope, the optimal multiplicity of infection, one-step growth curve and other biological characteristics experiments and whole genome sequencing analysis. A strain of E. coli phage vB_EcoM-ZQ1 was successfully isolated. The phage belonged to the order Ceratoviridae and the Mycoviridae family. The optimal multiplicity of infection was 0.1, the incubation period was 20 min, the lysis period was 10 min, and the amount of lysis was 92 PFU·cell^-1. Under the conditions of temperature 4-55℃ and pH4-10, the phage had high activity. It had a certain lysis effect on multi-drug resistant E. coli, with a lysis rate of 25.5%. The phage gene had a total length of 149 112 bp and a GC content of 49.1%. Among 184 ORFs, 70 ORFs encoded functional proteins, and 2 ORFs encoded tRNAs, and no drug resistance genes and virulence factors were found; The BLAST comparison found that the phage had 92.37% similarity with Shigella phage phiSboM-AG3. The results of a homologous phylogenetic tree constructed based on the large subunit of the phage terminal enzyme showed that the phage vB_EcoM-ZQ1 was closely related to the Salmonella phage pSal-SNUABM-02. The results indicated that the bacteriophage vB_EcoM-ZQ1 had stable biological characteristics and complete gene structure, which had laid a certain preliminary foundation for the follow-up prevention and control of bla_NDM-5 and mcr-1 positive E. coli.

Key words: phage, biological characteristics, Escherichia coli, bla_NDM-5, mcr-1

中图分类号:

Q939.48

刘畅, 刘教, 陈进, 王勉之, 熊文广, 曾振灵. 1株可裂解bla_NDM-5及mcr-1阳性大肠杆菌噬菌体的分离与生物学特性[J]. 畜牧兽医学报, 2022, 53(3): 857-866.

LIU Chang, LIU Jiao, CHEN Jin, WANG Mianzhi, XIONG Wenguang, ZENG Zhenling. Isolation and Biological Characteristics of a Bacteriophage Capable of Lysing bla_NDM-5 and mcr-1 Positive Escherichia coli[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 857-866.

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1株可裂解bla_NDM-5及mcr-1阳性大肠杆菌噬菌体的分离与生物学特性

Isolation and Biological Characteristics of a Bacteriophage Capable of Lysing bla_NDM-5 and mcr-1 Positive Escherichia coli

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1株可裂解blaNDM-5及mcr-1阳性大肠杆菌噬菌体的分离与生物学特性

Isolation and Biological Characteristics of a Bacteriophage Capable of Lysing blaNDM-5 and mcr-1 Positive Escherichia coli

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1株可裂解bla_NDM-5及mcr-1阳性大肠杆菌噬菌体的分离与生物学特性

Isolation and Biological Characteristics of a Bacteriophage Capable of Lysing bla_NDM-5 and mcr-1 Positive Escherichia coli