体外拮抗幽门螺杆菌抑菌方法的研究

目的: 建立和优化针对幽门螺杆菌的体外抑菌方法,为筛选拮抗幽门螺杆菌的菌株和活性物质提供简单有效的操作方法。方法: 将幽门螺杆菌NCTC 11637国际标准株作为指示菌,比较研究打孔扩散法、纸片法、点状覆盖法、共培养活菌计数法等常规抑菌方法,对各种方法在体外拮抗幽门螺杆菌实验的适用性、操作难度、判定时间等方面进行评价。结果: 纸片法样品装载量少,点状覆盖法无法检测,活菌计数法操作烦琐,批处理量小,而优化后的打孔扩散法具有简单高效,判定时间短,观察结果直观等特点。结论: 优化后的打孔扩散法符合幽门螺杆菌体外拮抗实验方法的要求,适用于多样品检测,为筛选拮抗幽门螺杆菌菌株和分离纯化活性物质过程中的各步骤间活性检测提供了极大的帮助。

Objective: To establish and optimize a new antibacterial method in vitro providing a simple and effective operation for selecting bacterial strains and active substance with function of anti-Helicobacter pylori. Methods: Using Helicobacter pylori NCTC 11637 as an indicator bacteria,compare the applicability,difficulties of operation and time requirement for determination among agar well diffusion assay,disc diffusion assay,dotted overlay assay and coculture experiment for living cell count assay. Results: Disc diffusion assay had low sample loading capacity;dotted overlay assay was difficult to detect and the operation process of living cell detection is complex,with small batch quantity.However,optimized well diffusion assay had three characteristics:it was simple and effective;time for determination was short and the results could be intuitively observed. Conclusion: Optimized well diffusion assay meets the requirement for in vitro antibacterial experiment,suitable for detection of a large number of samples,offering significant benefits for the intermediate step detection of bacterial strain selected and active substance isolated and purified with funciton of anti-Helicobacter pylori.

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