Department of Clinical Laboratory, Beijing Youan Hospital, Capital Medical University, Beijing, People's Republic of China.
General Surgical Center, Beijing Youan Hospital, Capital Medical University, Beijing, People's Republic of China.
Am J Trop Med Hyg. 2023 Apr 24;108(6):1220-1226. doi: 10.4269/ajtmh.22-0303. Print 2023 Jun 7.
This study aimed to isolate and characterize phages as an alternative treatment of multidrug- or pan-drug-resistant Pseudomonas aeruginosa. Phage titers and bacterial densities correlated, with the phages disappearing after bacteria were eliminated. We isolated phages in filtered sewage water by a double-layered agar spot test. Fifty-eight P. aeruginosa strains were used to screen the host spectrum of the 14 phages isolated. Random amplification of polymorphic DNA-typing polymerase chain reaction was used to analyze the genomic homologies of the 58 host bacteria strains and four phages with a broad host spectrum. Transmission electron microscopy was used to observe the morphology of the four phages with a broad host spectrum. Mice with intraabdominal P. aeruginosa infection were used as an in vivo animal model to investigate the therapeutic effect of the selected phage. Four virulent phages with a broad host spectrum specific to P. aeruginosa strains were isolated. They were all double-stranded DNA viruses and belonged to four different genotypes. The test curve showed that phage I had the highest adsorption rate, the shortest latent period, and the largest burst size. The infected mouse model indicated that small doses of phage I could prevent the death of infected mice. Phage titers and bacterial densities correlated, with phages disappearing after bacteria were eliminated. Phage I was the most effective and promising treatment of drug-resistant P. aeruginosa.
本研究旨在分离和鉴定噬菌体作为治疗多重耐药或泛耐药铜绿假单胞菌的替代方法。噬菌体效价和细菌密度相关,细菌被清除后噬菌体也消失了。我们通过双层琼脂点试验从过滤污水中分离噬菌体。用 14 株分离的噬菌体筛选 58 株铜绿假单胞菌,以确定其宿主范围。随机扩增多态性 DNA-聚合酶链反应用于分析具有广谱宿主的 58 株宿主细菌和 4 株噬菌体的基因组同源性。透射电子显微镜观察具有广谱宿主的 4 株噬菌体的形态。用腹腔内铜绿假单胞菌感染的小鼠作为体内动物模型,研究所选噬菌体的治疗效果。分离出 4 株具有广谱宿主特异性的毒力噬菌体,均为双链 DNA 病毒,属于 4 种不同基因型。测试曲线表明噬菌体 I 具有最高的吸附率、最短的潜伏期和最大的爆发量。感染小鼠模型表明,小剂量的噬菌体 I 可预防感染小鼠的死亡。噬菌体效价和细菌密度相关,细菌被清除后噬菌体也消失了。噬菌体 I 是治疗耐药铜绿假单胞菌最有效和最有前途的方法。