一种对铜绿假单胞菌具有活性的新型噬菌体PUTH1的分离与鉴定

Isolation and characterization of a novel bacteriophage PUTH1 active against Pseudomonas aeruginosa.

作者信息

Tan Qizhao, Ye Jing, Zhu Tengjiao, Tian Yun

机构信息

Department of Orthopaedics, Peking University Third Hospital, Beijing, 100191, China.

Engineering Research Center of Bone and Joint Precision Medicine, Ministry of Education, Beijing, 100191, China.

出版信息

Virol J. 2025 Jul 19;22(1):248. doi: 10.1186/s12985-025-02859-8.

DOI:10.1186/s12985-025-02859-8

PMID:40684166

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12275284/

Abstract

BACKGROUND

() is a leading cause of healthcare-associated infection and is prone to antibiotic resistance. Treating infections caused by antibiotic-resistant remains clinically challenging. The increasing prevalence of multidrug-resistant (MDR) bacteria has prompted the development of alternatives to antibiotics. Bacteriophage (phage) therapy has been regarded as a promising antibacterial alternative in many cases of MDR bacterial infections. This study aimed to isolate and characterize phages with therapeutic potential against MDR .

METHODS

Phage PUTH1 was isolated from filtered sewage water collected at Peking University Third Hospital using the double-layer agar spot method. The phage morphology was characterized using transmission electron microscopy (TEM). Subsequently, key biological properties were evaluated, including temperature and pH stability, one-step growth curve, adsorption rate, and optimal multiplicity of infection (MOI). Additionally, the anti-biofilm activity was assessed using crystal violet staining assays. Finally, whole genome sequencing was conducted, followed by comprehensive bioinformatics analysis.

RESULTS

Morphological and genomic characterization confirmed that the phage PUTH1 belongs to the Podoviridae family. The phage exhibits remarkable thermal stability (active up to 60 °C) and across a broad pH range (pH 4.0–9.0). Furthermore, the phage is characterized by a short latent period (approximately 30 min), a large burst size (54.5 PFU/cell), and rapid adsorption efficiency, achieving approximately 80% adsorption after 6 min. PUTH1 exhibits significant anti-biofilm activity. The genome of phage was 45,483 bp with 52.25% GC content and contained 70 putative genes; functions were predicted for 32 of them. Phylogenetic analysis revealed that PUTH1 is distinct from currently known phages and belongs to a novel species. The genome sequencing analysis suggested that it does not exist any known virulence and antibiotic resistant genes.

CONCLUSION

Therefore, our findings demonstrate that this novel phage represents a promising therapeutic candidate for treating MDR infections.

SUPPLEMENTARY INFORMATION

The online version contains supplementary material available at 10.1186/s12985-025-02859-8.

摘要

背景

（）是医疗保健相关感染的主要原因，且易于产生抗生素耐药性。治疗由抗生素耐药菌引起的感染在临床上仍然具有挑战性。多重耐药（MDR）细菌的日益流行促使人们开发抗生素的替代品。在许多MDR细菌感染病例中，噬菌体疗法被视为一种有前景的抗菌替代方法。本研究旨在分离和鉴定对MDR具有治疗潜力的噬菌体。

方法

采用双层琼脂斑点法从北京大学第三医院收集的过滤污水中分离出噬菌体PUTH1。使用透射电子显微镜（TEM）对噬菌体形态进行表征。随后，评估关键生物学特性，包括温度和pH稳定性、一步生长曲线、吸附率和最佳感染复数（MOI）。此外，使用结晶紫染色试验评估抗生物膜活性。最后，进行全基因组测序，随后进行全面的生物信息学分析。

结果

形态学和基因组特征证实噬菌体PUTH1属于短尾噬菌体科。该噬菌体表现出显著的热稳定性（在高达60°C时仍有活性），并且在较宽的pH范围内（pH 4.0 - 9.0）均有活性。此外，该噬菌体的特点是潜伏期短（约30分钟）、裂解量高（54.5 PFU/细胞）和吸附效率快，6分钟后吸附率达到约80%。PUTH1表现出显著的抗生物膜活性。噬菌体基因组为45,483 bp，GC含量为52.25%，包含70个推定基因；其中32个基因的功能得到了预测。系统发育分析表明PUTH1与目前已知的噬菌体不同，属于一个新物种。基因组测序分析表明它不存在任何已知的毒力和抗生素耐药基因。