分枝杆菌噬菌体生物学特性探讨

An approach to biological characters of mycobacteriophage

为了确定不同分枝杆菌噬菌体的宿主菌以及扩增方法和最佳保存方法,观察了七种分枝杆菌噬菌体对结核分枝杆菌、耻垢分枝杆菌的裂解情况,并分别于感染后 24、48小时采用离心 过滤、孵育 过滤方法收集噬菌体比较扩增效率,采用不同稳定剂对分枝杆菌噬菌体进行液体和冻干保存,在不同时间段采用琼脂双层法检测其效价。结果显示:①D29分枝杆菌噬菌体能同时较高效地裂解结核分枝杆菌和耻垢分枝杆菌;②感染 48小时后采用孵育 过滤方法收集的噬菌体效价高,方法简单;③液体 4℃保存的噬菌体稳定性好, 70℃液体保存和冻干后 4℃、室温、37℃保存依据不同稳定剂而相差较大。因此,在 48小时后采用孵育 过滤方法收集噬菌体具有高效率特性并且简单易行,噬菌体液体 4℃保存简单、有效,值得推荐。

To study the host mycobacteria of different mycobacteriophages and compare various conditions for replication and preservation of mycobacteriophage, the host mycobacteria of seven mycobacteriphages were confirmed, including M.tuberculosis and M.smegmatis. The titers of replicated mycobacteriphage with different methods in 24 or 48 hours were compared, including centrifugation-filtration method and incubation-filtration method with 0.2μm filter membrane. The mycobac-teriophage treated with different methods and stablizers were tested at different time by double-agar method. The results included: ①Phage D29 could highly lyse M.tuberculosis and M.smegmatis; ② The titer of replicated phage was higher when the host had been infected for 48 hours with the incubation-filtration method; ③The phage stored at 4℃ was more stable than that under the lyophilized condition, which is simple and efficient without using special equipment.