分枝杆菌噬菌体DNAⅢ生物学特性及其抗耐药结核潜力  

Biological characteristics of mycobacteriophage DNAⅢ and the potentials in control of drug-resistant tuberculosis

作　　者：邬亭亭[1,2] 刘平 郭术良 魏强 罗永艾[2] WU Tingting;LIU Ping;GUO Shuliang;WEI Qiang;LUO Yong＇ai(Department of Respiratory Medicine,The Third People＇s Hospital of Chengdu,Chengdu G10031,China;Department of Respiratory and Critical Care Medicine,The First Affiliated Hospital of Chongqing Medical University,Chon g qin g 40 O01 G,China;Department of Respiratory Medicine,The People＇s Hospital of Changshou,Chongqing 401220,China;Department of Internal Medicine,Chengdu Tian f u New District People＇s Hospital,Chengdu 610213,China)

机构地区：[1]成都市第三人民医院呼吸内科,四川成都610031 [2]重庆医科大学附属第一医院呼吸与危重症医学科,重庆400016 [3]重庆市长寿区人民医院呼吸内科,重庆401220 [4]成都市天府新区人民医院内二科,四川成都610213

出　　处：《西部医学》2018年第9期1264-1268,共5页Medical Journal of West China

基　　金：国家“十二五”重大专项课题(2012ZX10003009);四川省卫计委科研课题(17PJ381)

摘　　要：目的研究分枝杆菌噬菌体DNAⅢ的生物学特性及抗耐结核潜力,筛选鸡尾酒疗法配方噬菌体。方法采用双层琼脂培养法制备噬菌体DNAⅢ,观察噬菌斑特点;透射电镜观察DNAⅢ形态;噬菌斑实验检测DNAⅢ最佳和最小感染复数(MOI);通过一步生长曲线实验确定DNAⅢ潜伏期及裂解量;检测DNAⅢ对温度、酸碱度的耐受情况;检测DNAⅢ对分枝杆菌的裂解谱;血清中和实验检测DNAⅢ的抗原性,及其与其它8种分枝杆菌噬菌体之间的亲缘关系。结果 DNAⅢ噬菌斑圆形透明,直径约1mm;DNAⅢ尾长(208±28.7)nm;最佳MOI为10-4,DNAⅢ对耻垢分枝杆菌极度易感;DNAⅢ感染宿主菌的潜伏期约为165min,裂解量为28;DNAⅢ对热、酸碱均敏感;DNAⅢ能裂解耻垢分枝杆菌、结核分支杆菌标准株、多数结核分枝杆菌临床耐药株;DNAⅢK值为770.01,抗血清对非对应噬菌体的中和活性差。结论 DNAⅢ抗原性低,裂解谱广,具有抗耐药结核作用,可作为鸡尾酒疗法的候选噬菌体。Objective To explore the biological characteristics of phage DNAⅢ and its anti-drug-resistant tuberculosis potentials.Methods We observed morphological property of the plaques by double-layer plating and ltra-structure of DNAⅢ by electron microscopy.We examined DNAⅢ＇s optimal muitiplicity of infection and its single step growth curve and inspected how DNAⅢ＇s survived was affected by temperature,pH value.We determined the host range of phage by Spot Test and the reaction constants between DNAⅢ antiserum and 33 D,D29,Legendre,Leo,DNAⅢ,TM4,Bo4,Sedeg and Clark by neutralization test.Results The plaques of DNAⅢ were transparent and their sizes were about 1 mm in diameter.DNAⅢ had a long tail（208±28.7 nm in length）.10-4 MOI infected host bacteria gave the highest phage off springs.0 ur single step growth test showed that phage DNAⅢ had a latent period of 165 minutes and a burst size of 28 and it was sensitive to temperature and pH value.The susceptibility to phage DNAⅢ was also investigated with other clinical strains of drug-resistant Mycobacterium tuberculosis.K value between antiserum and DNAⅢis770.01 and there was little correlation among them.Conclusion Phage DNAⅢis a suitable broad host range and low antigenicity siphoviridae phage which could be used in phage cocktail therapy.

关 键 词：分枝杆菌噬菌体DNAⅢ 生物学特性 耐药结核分枝杆菌 

分 类 号：R52[医药卫生—内科学]