一种呼吸道口咽菌群系统分类和快速鉴定的方法

A method for systematic classification and rapid identification of oropharyngeal microbiota

目的:探索有效鉴别呼吸道口咽部菌群的方法。方法:取108例健康青年志愿者咽拭子标本,进行传统细菌培养,定性、定量分析培养菌落,对特征菌落PCR扩增16SrDNA片段后测序,并与NCBI基因序列库比对。结果:完成了全部108例标本口咽菌群在4种细菌培养基、1种真菌培养基上生长的27种菌落的特征分类。菌落PCR有效扩增出细菌特征菌落的16SrDNA片段,经序列分析后在属、种水平鉴定细菌。结论:初步建立了一种呼吸道口咽菌群系统分类和快速鉴定的方法。该方法是通过分离培养口咽菌群并对特征菌落扩增其16SrDNA片段,再进行序列分析鉴定细菌,为有效评价口咽菌群的健康状态提供直接依据。

Objective: To explore an effective method for identifying oropharyngeal microbiota. Methods: Pharyngeal swab specimens from 108 healthy young volunteers were collected and subjected to bacterial culture. Colonies were analyzed qualitatively and quantitatively. The 16S rDNA fragments of characteristic colonies were amplified and sequenced, and the sequencing results were compared with Genbank Entries(BLAST). Results: The characteristics of 27 colonies of oropharyngeal microbiota in 108 specimens grown on 4 kinds of bacterial and 1 kind of fungal media were classified. The 16S rDNA fragments of bacterial characteristic colonies were effectively amplified by colony PCR. The bacteria were identified at genus and species level after sequence analysis. Conclusions: A method of systematic taxonomy and rapid identification of respiratory oropharyngeal microbiota is preliminarily established. The method can be used to culture oropharyngeal microbiota and amplify 16S rDNA fragments of characteristic colonies, and then identify bacteria by sequence analysis. This method provides a direct basis for the effective evaluation of the health status of oropharyngeal microbiota.