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目的:探讨微量点样法用于计数微生物活菌数的可行性。方法:分析了菌种、培养基和点样量对测定结果的影响,并与平板涂布法计数结果进行了比较和相关性及线性回归分析。结果:微量点样法的菌落生长特点受菌种、培养基和点样量的影响较小;活菌计数结果与平板涂布法相比,(1)对8种测试菌株(约108CFU/mL)均没有显著性差异(P>0.05);(2)对4种不同类型细菌(Escherichia coli,Pseudomonas aeruginosa,Streptococcus pneumoniae and Bacteroides fragilis,约107～109CFU/mL)均具有极显著的直线回归关系(P<0.01,n=5),其回归方程分别为y=1.0740x-0.4144(R2=0.9881),y=0.8100x+1.4789(R2=0.9977),y=1.1194x-0.7373(R2=0.9806),y=0.9107x+0.772(R2=0.9812)。结论:微量点样法简化了平板涂布法的操作过程,适应性广、准确性高,因此,是一种简单、经济的细菌活菌计数方法。
Objective: To explore the feasibility of using micro-spotting method for counting viable microorganisms. Methods: The effects of bacteria, culture medium and sample volume on the determination results were analyzed. The results were compared with the flat coating method and the linear regression analysis. Results: The colony growth characteristics of the micro-spotting method were less affected by the strain, medium and sample volume. Compared with the flat-plate coating method, (P> 0.05). (2) There was a significant linear regression between four different types of bacteria (Escherichia coli, Pseudomonas aeruginosa, Streptococcus pneumoniae and Bacteroides fragilis, about 107-109 CFU / mL) (R2 = 0.9977), y = 1.1194x-0.7373 (R2 = 0.9806), y = 0.8100x + 0.4789 (R2 = = 0.9107x + 0.772 (R2 = 0.9812). Conclusion: The micro-spotting method simplifies the operation of the plate coating method, wide adaptability and high accuracy, therefore, is a simple and economical method of counting bacterial viable bacteria.