快速从医院废水中大规模分离铜绿假单胞杆菌噬菌体  

A Fast Method for Large-Scale Isolation of Pseudomonas aerugino sa Phage from the Hospital Sewage

作　　者：范华昊[1,2] 范俊芬[2] 滑玉会[2] 裴广倩[2] 张文惠[2] 张博[2] 陈晨[2] 李建彬[2] 王晓娜[2] 安小平[2] 米志强[2] 王琳[3] 尹秀云[3] 陈建魁[3] 童贻刚[1,2] 

机构地区：[1]中南大学基础医学院,湖南长沙410013 [2]军事医学科学院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京100071 [3]军事医学科学院附属医院临床检验科,北京100071

出　　处：《生物技术通讯》2012年第4期567-571,共5页Letters in Biotechnology

基　　金：国家自然科学基金(81072350;30872223);国家高技术研究发展计划(2009AA02Z111);重大新药创制科技重大专项"十二五"实施计划(2011ZX09401-023);病原微生物生物安全国家重点实验室开放课题(PBS2009A-04)

摘　　要：目的:从医院废水中快速分离多株不同的铜绿假单胞杆菌噬菌体,研究其生物学特性,为建立铜绿假单胞杆菌噬菌体库做准备。方法:利用噬菌斑法从未经处理的医院污水中分离和鉴定铜绿假单胞杆菌噬菌体,根据感染谱的不同确定它们为不同的铜绿假单胞杆菌噬菌体;重点研究其中一株宿主谱较广的噬菌体的生物学特性,采用负染法电镜观察噬菌体的形态和大小,提取该噬菌体的基因组并进行酶切电泳分析,测定噬菌体感染复数并观察其一步生长曲线。结果:通过噬菌斑法分离出90株铜绿假单胞杆菌噬菌体。电镜观察显示,噬菌体Pa27P1头部呈立体对称,有一长尾;酶切结果显示,噬菌体Pa27P1的基因组为双链DNA;生长曲线表明噬菌体Pa27P1感染宿主菌的潜伏期为25 min,爆发时间为25 min,裂解量为514。结论:90株铜绿假单胞杆菌噬菌体中有5株具有较广的噬菌谱,其组合能裂解所有18株铜绿假单胞杆菌,为深入研究铜绿假单胞杆菌噬菌体的生物学特性及其功能提供了依据。Objective： To investigate biological properties of Pseudomonas aeruginosa phages rapidly isolated from hospital raw sewage, and lay the foundation of establishment of P.aeruginosa phage library. Methods： According to the different spectrum of infection, different bacteriophages were identified using plaque methods, and some biological characteristics of a bacteriophage Pa27P1 which has a wide host spectrum were studied. The electron micrographics of phages were carried out to obverse the morphology and size of the bacteriophage by negative staining. The extraction of phage genome and nuclease digestion of its genomic DNA was performed. The multiplicity of optimal infection and one-step growth curve of the phage were determined. Results： Ninety P.aeruginosa phages were identified. Electron micrographics showed that bacteriophage Pa27P1 exhibited a cubic symmetry, and this phage had a long tail. The growth curve showed that the incubation period was 25 minutes, the time of burst phase was 25 min, and the burst size was 514. Conclusion： Five strains of the phages have a broader host range in P.aeruginosa, and their mixture can crack all these eighteen strains. These data enable us to explore more properties of P.aeruginosa phages.

关 键 词：铜绿假单胞杆菌噬菌体 大规模分离 细菌耐药 生物学特征 

分 类 号：Q939.48[生物学—微生物学] R373[医药卫生—病原生物学]