药品微生物实验室水系统分离37株微生物的鉴定及分析

Identification and Analysis of 37 Strains Isolated from Water System of Pharmaceutical Microbiology Laboratory

目的针对目前国内微生物实验室试验用水的实际情况,探索试验用水微生物群落的组成及分布规律。方法参考国内外药典对纯化水微生物的检验方法,收集某实验室7个纯水使用点的纯化水,经薄膜过滤法处理,采用R2A琼脂培养基培养、分离和纯化,并进行革兰染色。分别运用基因型鉴定方法(16S rDNA序列分析、Riboprinter全自动微生物基因指纹鉴定系统),生化鉴定方法(Biolog自动微生物鉴定系统)对检出微生物进行鉴定,并对实验结果进行分析比较。结果本研究共培养获得37株细菌,主要为鞘氨醇单胞菌属和叶杆菌属等。16S rDNA序列分析能对37株微生物达到属或属水平以上的鉴定,Biolog自动微生物鉴定系统鉴定出13株微生物,Riboprinter全自动微生物基因指纹鉴定系统鉴定出17株微生物。结论16S rDNA序列分析适合对水系统中的微生物进行基于培养的群落分布分析,Riboprinter全自动微生物基因指纹鉴定系统更适用于同种/属细菌的分型及溯源分析。

Objective To explore the composition and distribution law of microbial community in test water according to the actual situation of test water system of domestic microbial laboratory.Methods According to microbiological test method for purified water listed in the domestic and foreign pharmacopoeia,the purified water samples from seven pure water sites in the laboratory were collected,then were treated by membrane filtration,incubated on the R2A agar medium,and stained by Gram staining after isolation and purification.The micrograms were identified by 16S rDNA gene sequence analysis,Riboprinter automatic microbial genetic fingerprint identification system and physiological and biochemical methods(Biolog automated microbes identification system),and the results were analyzed and compared.Results A total of 37 strains microbes were cultured and obtained in the study.The composition of microorganisms in water system were mainly Sphingomonas,Phyllobacterium and so on.37 strains of microorganisms at or above the genus level were identified by 16S rDNA sequence analysis.13 and 17 strains were identified by Biolog and Riboprinter,respectively.Conclusion 16S rDNA sequence analysis is suitable for culture based community distribution analysis of microorganisms in water system,and Riboprinter automatic microbial genetic fingerprint identification system is more suitable for the classification and traceability analysis of same species/genus.