摘要
目的从地下污水中分离出一株鲍曼不动杆菌噬菌体并对其进行研究,为噬菌体治疗鲍曼不动杆菌感染提供理论基础。方法以鲍曼不动杆菌GY-6为宿主菌,采用双层琼脂平板法分离噬菌体,对其进行生物学特性检测、治疗效果评价及遗传信息分析。结果分离到鲍曼不动杆菌噬菌体Abgy202162。透射电镜(TEM)分析显示,Abgy202162的形态呈二十面体结构。生物学特性分析表明其最佳感染复数为1,潜伏期为5 min,暴发量大小约为每个细胞520 PFU。此外,Abgy202162在不同浓度的三氯甲烷以及不同pH水平和温度下保持稳定。经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析,其含有10种蛋白质,分子量为15~100 ku。Abgy202162的双链DNA基因组由40889 bp组成,G+C含量为38.85%,包含47个开放阅读框(ORFs),其中26个ORFs具有特异性功能,未发现毒力相关基因或抗生素耐药基因。系统发育分析显示,Abgy202162是自转录噬菌体科,拜耶林克噬菌体亚科,弗留纳病毒属的一个新的噬菌体。Abgy202162在大蜡螟幼虫体内模型中表现出了治疗鲍曼不动杆菌感染的能力。结论噬菌体Abgy202162对环境耐受性强、安全性高,提示其有望成为抗生素的代替治疗剂用于治疗鲍曼不动杆菌引起的感染。
Objective To isolate a Acinetobacter baumannii(Ab)phage from underground sewage,study its properties,and to provide a theoretical basis for phage treatment of Ab infection.Methods Double-layer agar technique was used to isolate phages by using Ab GY-6 as the host strain.Biological characterization and therapeutic effect of the phage was tested.Genetic information of the phage was analyzed.Results Ab phage Abgy202162 was isolated.Transmission electron microscopy(TEM)analysis showed that the morphology of Abgy202162 exhibited an icosahedral structure.Biological characteristic analysis showed that the optimal multiplicity of infection was 1,the latent period was 5 min,and the burst size was approximately 520 PFU per cell.In addition,Abgy202162 remained stable at different concentrations of chloroform,pH,and temperatures.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)analysis showed that it contained 10 proteins with molecular weights ranging from 15 to 100 ku.The double-stranded(ds)DNA genome of Abgy202162 consisted of 40889 bp and its G+C content was 38.85%.It contained 47 open reading frames(ORFs),of which 26 had specific functions,but no virulence related genes or antibiotic resistance genes were found.Phylogenetic analysis showed that Abgy202162 was a new phage in the Autographiviridae family,Beijerinkvirinae subfamily,and Friunavirus genus.Abgy202162 showed the ability to prevent Ab infection in the Galleria mellonella in vivo model.Conclusion The phage Abgy202162 has strong environmental tolerance and high safety,indicating its potential as an antibiotic alternative used in the treatment of infections caused by Ab.
作者
田询
谭文才
杨碧
刘翔
禹文峰
齐晓岚
江银辉
Tian Xun;Tan Wencai;Yang Bi;Liu Xiang;Yu Wenfeng;Qi Xiaolan;Jiang Yinhui(Key Laboratory of Medical Molecular Biology of Guizhou Province,Guizhou Medical University,Guiyang 550004;Guiyang&Key Laboratory of Endemic and Ethnic Diseases,Ministry of Education,Guizhou Medical University,Guiyang 550004)
出处
《安徽医科大学学报》
CAS
北大核心
2024年第10期1742-1751,共10页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金项目(编号:32000017、31660012)。
关键词
鲍曼不动杆菌
噬菌体
生物学特性
基因组分析
治疗效果评价
大蜡螟幼虫
Acinetobacter baumannii
phage
biological characteristics
genomic analysis
therapeutic evaluation
Galleria mellonella larvae
