幽门螺杆菌悉尼标准菌株传代培养方法研究

Study on Subculture Methods of Helicobacter pylori Sydney-Standard Strain

目的:比较不同培养方法幽门螺杆菌(H.pylori)悉尼标准菌株(SS1菌株)的生长速度,寻找简便、快捷的H.pylori培养方法。方法:通过4种方法对SS1菌株进行培养,在哥伦比亚血平板中加入脑心浸润液及菌液,使用无菌一次性接种环将菌液均匀涂布。方法一是将完成菌液涂布的哥伦比亚血平板放入装有厌氧产气包的密封厌氧袋中;方法二是将完成菌液涂布的哥伦比亚血平板置于含去离子水和厌氧产气包的密封培养罐中;方法三是在方法二的基础上,在密封培养罐内点燃1次蜡烛;方法四是在方法三的基础上,于密封培养罐内多次点燃蜡烛。4种方法均置于CO_(2)培养箱内进行培养,通过观察菌落形态、革兰染色和质谱检测鉴定H.pylori菌株。结果:4种方法均能成功培养出H.pylori菌株,在达到同等菌落数量情况下,方法四用时显著少于其他3种方法,差异有统计学意义(P<0.05)。结论:找到一种操作简单、价格低廉的H.pylori传代培养方法,具有较强的应用前景,值得推广。

Objective:To compare the growth rate of Helicobacter pylori(H.pylori)Sydney-standard strain(SS1 strain)by different culture methods,and to find a simple and fast culture method for H.pylori.Methods:The SS1 strain was cultured by four methods.Brain-heart infiltration fluid and bacterial fluid were added to Columbia blood plate,and the bacterial fluid was evenly spread with a disposable inoculating loop.Method 1 was to put the Columbia blood plate coated with bacterial liquid into a sealed anaerobic bag equipped with an anaerobic gas production bag.Method 2 was to place the Columbia blood plate coated with the bacterial solution in a sealed culture jar containing deionized water and an anaerobic gas production bag.Method 3 was based on method 2,in which a candle was lit once in the sealed culture jar.Method 4 was based on method 3,in which a candle was lit in the sealed culture jar for many times.The four methods were all completed in a CO_(2)incubator,and the H.pylori strains were identified by observing colony morphology and Gram staining,and detected by mass spectrometry.Results:H.pylori strains were successfully cultivated by the four methods.When the number of colonies reached the same level,Method 4 takes significantly less time than the other three methods,and the differences were statistically significant(P<0.05).Conclusion:A simple and inexpensive method of H.pylori subculture has been found,which has strong application prospects and is worthy of promotion.