鲍曼不动杆菌噬菌体的分离及基因组分析

Isolation and genomic analysis of the phage of Acinetobacter baumanni

试验以耐药性鲍曼不动杆菌为宿主菌,采取双层琼脂平板法,从污水中分离纯化鲍曼不动杆菌噬菌体,并筛选裂解谱相对较宽的噬菌体,测定其最佳感染复数(MOI),建立一步生长曲线;扩增、浓缩噬菌体颗粒,应用透射电镜观察噬菌体形态,提取、鉴定其基因组类型;以期为丰富鲍曼不动杆菌的噬菌体库、临床中相关疾病的诊疗提供参考。结果显示,经纯化筛选得到一株宽裂解谱、强裂解活性的噬菌体,命名为ABC-Hγ。该噬菌体MOI为0.01,潜伏期约为30 min,爆发期持续时间较长,暂分类为有尾病毒目肌尾病毒科,电泳鉴定其为双链DNA病毒。研究表明,噬菌体ABC-Hγ在体外具有高效的杀菌活性及生物学稳定性,在治疗鲍曼不动杆菌引起的动物疫病方面具有较大的应用潜力。

The bacteriophage of Acinetobacter baumannii was isolated and purified from wastewater by double-layer AGAR plate method,and the bacteriophage with relatively wide cleavage spectrum was screened,the optimal number of infection(MOI)was determined,and the one-step growth curve was established.The phage particles were amplified and concentrated,the morphology of phage was observed by transmission electron microscopy,and the genomic types were extracted and identified.In order to provide reference for enriching the phage library of Acinetobacterbaumannii and the diagnosis and treatment of related diseases in clinic.The results showed that a phage with wide cleavage spectrum and strong cleavage activity was obtained after purification and screening,and was named ABC-Hγ.The MOI of the bacteriophage was 0.01,the incubation period was about 30 min,and the outbreak lasted for a long time.The bacteriophage was temporarily classified as Myoviridae in the order Caudovirales,and was identified as a double-stranded DNA virus by electrophoresis.Studies have shown that phage ABC-Hγhas high bactericidal activity and biological stability invitro,and has great application potential in the treatment of animal diseases caused by Acinetobacterbaumannii.