产超广谱β-内酰胺酶大肠埃希菌噬菌体分离及其生物学特性观察  

Biological characteristics investigation and the isolation of extended-spectrum β-lactamases-producing Escherichia coli Phage with broad host range

作　　者：徐敏超[1] 胡北[1] 严群[1] 刘双又[1] 胡俊波[1] 朱旭慧[2] 孙自镛[2] 吴乙璇[3] 杨威[3] 王晶[3] 

机构地区：[1]华中科技大学同济医学院附属同济医院分子医学中心,湖北武汉430030 [2]华中科技大学同济医学院同济医院检验科,湖北武汉430030 [3]华中科技大学同济医学院免疫学研究所,湖北武汉430030

出　　处：《中国现代医学杂志》2010年第22期3368-3371,共4页China Journal of Modern Medicine

基　　金：武汉市科委"青年科技晨光计划"(No:20025001026)

摘　　要：目的分离、筛选宽噬产超广谱β-内酰胺酶(ESBLs)大肠埃希菌噬菌体,观察其生物学特性。方法以30株产ESBLs大肠埃希菌为宿主菌,从医院污水中分离获得噬菌体,筛选出宽噬噬菌体,进行吸附实验、一步生长实验、噬菌体DNA初步酶切分析和电镜观察。结果所筛选出的宽噬噬菌体φ9882其宽噬率为36.7%,滴度为8.1×1018pfu/mL。5分钟吸附率约为98%,潜伏期约为30～40min,裂解量约为110。φ9882的DNA约为30kb。电镜显示,φ9882具有直径70nm微球形颗粒头部,棱角不明显,可见100nm尾轴。结论分离、筛选所获得的φ9882为宽噬噬菌体,其裂解性较强,裂解谱宽,吸收率高,潜伏期短,可进一步用于相应耐药菌感染的治疗研究。【Objective】To investigate the biological characteristics of an extended -spectrum beta -lactamases （ESBLs） -producing Escherichia coli bacteriophage with broad host range isolated from hospital sewage.【Methods】 30 ESBL-producing E.coli strains were used to isolate specific phages from hospital sewage and then the phage with broad host range was screened.The phage was identified by adsorption rate,one step growth experiment and negative staining for transmission electron microscope （TEM）.【Results】The broad-host-range phage φ9882 was screened out and it could cause 36.7% bacteriolysis among all those 30 ESBL-producing E.coli strains.About 98% of phage was absorbed to host bacteria within 5 minitues.The latent period is about 30~40 min and the burst size is about 110.The ultrastructure analysis under electron microscope demonstrated that the phage φ9882 has a roundshaped head with diameter 70 nm,and a short tail of 100 nm in length.【Conclusions】The phage φ9882 screened had relatively a broad host range,a high adsorption rate and a short latent period.The investigation of the its characteristics will provide the first experimental evidence to validate the clinical application of phage therapy.

关 键 词：大肠埃希菌 超广谱Β-内酰胺酶 耐药性 噬菌体 生物学特性 

分 类 号：R37[医药卫生—病原生物学]