基于培养组学的健康人口腔细菌分离鉴定初探

Isolation and identification of oral bacteria from healthy people based on cultureomics technology

目的 通过对口腔的不同部位进行细菌分离,扩展对口腔微生物多样性的认识,以期为后续的口腔菌群研究提供参考。方法 通过培养组学方法分离口腔不同部位的细菌,采集2个健康人扁桃体、唾液、牙菌斑3个部位的样本,使用13种培养基并于需氧和厌氧条件下进行培养,采用16S rRNA基因全长测序分析。结果 总计获得144株菌,分属36个种,10个科,3个门。其中,链球菌属细菌种类最多。基于16S rRNA基因全长相似度98.65%的阈值判定,获得5种潜在全新细菌。3个部位中,扁桃体分离的细菌种类多样性最丰富。结论 13种培养基中,BHI培养基获得的口腔细菌多样性最好;BAB培养基与TSA培养基更适合口腔链球菌生长,链球菌的比例更高;LBB培养基上只有较单一的口腔细菌生长,不适于广谱分离口腔细菌。

Objective To isolate bacteria from different parts of oral cavity, expand the understanding of oral microbial diversity and provide reference for subsequent research on oral microbial community. Methods Cultureomics technology was used to isolate bacteria collected from different oral parts including tonsil, saliva and dental plaque of two healthy volunteers. Thirteen media were used for cultivation under aerobic or anaerobic condition. The strains were identified based on 16 S rRNA gene sequencing. Results A total of 144 strains were isolated, which contained 5 potential new species(16 S rRNA gene similarity value <98.65%), distributed in 36 families, 10 classes, and 3 phyla. Among them, Streptococcus had the highest species richness. Among the three oral sampling sites, tonsil was seen to have the highest bacterial diversity. Conclusion Among the 13 media, BHI medium can result in a highest bacterial diversity. BAB and TSA media are suitable for Streptococcal growth. Only a few kinds of bacteria can grow on LBB medium, therefore it is not suitable for large-scale isolation of oral bacteria.