铜绿假单胞菌噬菌体PA-27-1的分离及其生物学特性

吴柳; 罗娟; 杨竹兰; 张波

doi:10.16016/j.1000-5404.201811044

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铜绿假单胞菌噬菌体PA-27-1的分离及其生物学特性

基础医学 | 更新时间：2026-01-09

- 铜绿假单胞菌噬菌体PA-27-1的分离及其生物学特性
- Isolation and biological characteristics of Pseudomonas aeruginosa phage PA-27-1
- 第三军医大学学报 2019年41卷第4期页码：315-322
- 作者机构：
  
  400038 重庆，陆军军医大学(第三军医大学)第一附属医院感染管理科
- 作者简介：
  
  [ 张波，E-mail：zhangbocq@aliyun.com ZHANG Bo, E-mail: zhangbocq@aliyun.com
- 基金信息：
  
  重庆市社会民生科技创新专项(cstc2015shmszx120110);军事医学临床重大领域陆军火箭军医疗技术创新计划项目(SWH2017ZDCX2009)
- DOI：10.16016/j.1000-5404.201811044
  中图分类号： R372;R393.9;R378.99
- 收稿：2018-11-07，
  
  修回：2018-12-07，
  
  纸质出版：2019-02-28
- 稿件说明：
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吴柳, 罗娟, 杨竹兰, 等. 铜绿假单胞菌噬菌体PA-27-1的分离及其生物学特性[J]. 第三军医大学学报, 2019,41(4):315-322.

Liu WU, Juan LUO, Zhulan YANG, et al. Isolation and biological characteristics of Pseudomonas aeruginosa phage PA-27-1[J]. Journal of Third Military Medical University, 2019, 41(4): 315-322.
吴柳, 罗娟, 杨竹兰, 等. 铜绿假单胞菌噬菌体PA-27-1的分离及其生物学特性[J]. 第三军医大学学报, 2019,41(4):315-322. DOI： 10.16016/j.1000-5404.201811044.

Liu WU, Juan LUO, Zhulan YANG, et al. Isolation and biological characteristics of Pseudomonas aeruginosa phage PA-27-1[J]. Journal of Third Military Medical University, 2019, 41(4): 315-322. DOI： 10.16016/j.1000-5404.201811044.

摘要

目的

从医院污水中分离筛选铜绿假单胞菌噬菌体，检测其裂解谱和生物学特性，探讨其应用价值。

方法

以铜绿假单胞菌标准菌株ATCC 27853和临床分离菌株为宿主，从未处理的医院污水中分离筛选铜绿假单胞菌噬菌体，检测其裂解谱，观测裂解效率最高的噬菌体的形态及生物学特性等。

结果

从污水中分离纯化到6株铜绿假单胞菌噬菌体，对162株不同药敏试验结果的临床菌株(包括63株多重耐药菌)裂解效率最高(79.63%)的噬菌体命名为PA-27-1，能裂解80.95%(51/63)的耐药菌株。噬菌体PA-27-1在双层琼脂平板上形成直径2~3 mm的圆形透明且边界清晰的噬菌斑；头部呈多面立体对称结构，尾短；最佳感染复数为0.000 1，与宿主菌液混合后即有80%的噬菌体吸附，孵育4 min后有98%的噬菌体吸附；潜伏期为0~20 min，以MOI=10试验时，裂解期为20~60 min，60 min后为平台期，裂解量约为248；在pH 6~10和≤50 ℃的环境中稳定性良好，对氯仿具有较高的耐受性，核酸类型为dsDNA。

结论

分离筛选到的铜绿假单胞菌噬菌体PA-27-1具有较宽的宿主谱，对多重耐药铜绿假单胞菌具有较强的裂解能力，吸附力强，裂解量大，酸碱和热稳定性好，可作为储备噬菌体，并为临床治疗铜绿假单胞菌感染提供基础和依据。

Abstract

Objective

To isolate and screen

Pseudomonas aeruginosa

(

P. aeruginosa

) phages from hospital sewage

identify their lysis spectra and biological characteristics

and explore their application value.

Methods

P. aeruginosa

standard strain ATCC 27853 and the clinical isolates in our department were used as hosts to isolate and screen

P. aeruginosa

phages from the untreated hospital sewage of our hospital. Then the lysis spectra of the obtained phages were examined. The morphological and biological characteristics of the phage of highest lysis efficiency was studied.

Results

Six

P. aeruginosa

phages were isolated and purified from the sewage. The phage with the highest lysis efficiency (79.63%) to 162 clinical isolates with different drug susceptibilities (including 63 multi-drug resistant strains) was named PA-27-1

and the phage lysed 80.95% (51/63) resistant strains. The plaque of the phage PA-27-1 was circular

transparent

and about 2~3 mm in diameter with clear borders

on a double-layer agar plate. Its head was multi-faceted and symmetrical

its tail was short

and the optimal multiplicity of infection was 0.000 1. Eighty percent of the phage was adhered to the host after being mixed with the host bacterial solution

and the percentage was increased to 98% after incubation for 4 min. One-step growth curve of phage PA-27-1 showed that its latent period were 0 to 20 min

the burst period was 20 to 60 min

and the platform period was subsequently

and the average burst size was about 248 (tested on MOI=10). It was stable in the environment of pH 6~pH 10 and ≤50 ℃

and had high tolerance to chloroform. Its nucleic acid type was dsDNA.

Conclusion

P. aeruginosa

phage PA-27-1 has a broad host spectrum

strong lysis ability to multi-drug resistant strains

large burst size

good acid-base and thermal stability. It could be used as a reserve phage and provide a scientific basis for the clinical treatment of

P. aeruginosa

infection.

关键词

Keywords

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