摘要
目的研究分枝杆菌噬菌体33D的生物学特性并探索其抗耐药结核的潜力。方法双层平板培养法制备噬菌斑,观察其特点;电镜观察33D形态;测定33D最佳感染复数和一步生长曲线;检测33D对紫外线、温度、酸碱度、酒精、氯仿的耐受性;斑点法测定33D的裂解谱;中和实验测定33D的吸附常数酶切分析确定33D核酸类型。结果33D噬菌斑圆形透明,边界清楚;33D尾长201nm,最佳MOI为10-5;33D感染宿主菌的潜伏期约为150min,裂解量为24;33D对紫外线、酸碱、酒精、氯仿、温度敏感;33D能裂解多数分枝杆菌临床耐药株;33D抗血清反应吸附常数K值为703.80;33D核酸为双链DNA分子。结论 33D属于长尾噬菌体科(siphoviridae),抗原性较低,裂解谱广,具有抗耐药结核潜力。
Objective We researched on the biological characteristics of phage 33D to explore its anti-drug-resistant tuberculosis potentials. Methods We observed morphological property of the plaques by double-layer plating and ultra- structure of 33D by electron microscopy. We examined 33D＇s optimal multiplicity of infection and its single step growth curve. And also we inspected how 33Ds survived was affected by UV ray, pH value, chloroform, temperature and alco- hol. We determined the host range of phage by Spot Test and the reaction constants between antiserum and 33D, D29, Legendre, Leo, DNAⅢ, TM4, BO4, Sedeg, Clark by neutralization test. We identified the type of nucleic acid of phage by restriction enzyme analysis. Results The plaques of 33D were transparent and their sizes were about 2 mm in diame- ter. 33D had a long tail about 201 nm in length. 0. 000 01 MOI infected host bacteria gave the highest phage offsprings. Our single step growth test showed that phage 33D had a latent period of 150 minutes and a burst size of 24 and it was sensitive to UV ray, pH value, chloroform, temperature and alcohol. The susceptibility to phage 33D was also investiga- ted with other clinical strains of Mycobacterium. Rate constant of reaction between antiserum and 33D is 703.80 and se- rological cross-neutralization test showed that there was little correlation among them. Restriction analysis indicated that phage 33D was a dsDNA virus with an approximate genome size of 43.2 kb. Conclusion Phage 33D is a suitable broad host range and low antigenicity siphoviridae phage which could be used in phage cocktail therapy.
出处
《西部医学》
2015年第11期1620-1625,共6页
Medical Journal of West China
基金
国家"十一五"科技重大专项(2008ZX10003-016)
关键词
分枝杆菌
噬菌体
生物学
形态学
Mycobacterium
Bacteriophage
Biology
Morphology
