基于不同通用引物比较分析肠道微生物群落变化

Comparison of intestinal microbial community succession based on different universal primer sets

肠道微生物对于人体健康的重要作用已经得到广泛证实,目前,对肠道微生物的研究大多采用基于扩增细菌16S rRNA基因V3-V4区的高通量测序分析,对古菌的关注较少。本研究选择了一对可以同时扩增细菌和古菌16S rRNA基因的引物,通过比较人为干扰肠道微生物前后的群落变化,说明这对引物适宜分析人类肠道细菌和古菌群落变化并具有一定优越性。采集志愿者粪便样品,同时用仅能扩增细菌引物(B引物)和细菌古菌通用引物(AB引物)进行扩增和高通量测序;使用几个常用的rRNA数据库判断引物对细菌的覆盖度和对古菌的扩增能力。结果表明,AB引物在可以展示B引物扩增出的细菌群落的基础上,可以得到肠道中常见的产甲烷古菌的序列,同时也展示出人为干扰肠道微生物前后的群落结构变化。AB引物可以仅通过一次扩增和测序同时分析肠道中的细菌和古菌群落,更加全面展示肠道微生物群落结构,适用于肠道微生物相关研究。

The important role of intestinal microorganisms in human health has been widely confirmed.At present,most of the studies on intestinal microorganisms are based on amplification of the V3-V4 region of bacterial 16S rRNA gene,and little attention has been paid to archaea.In this study,a primer set which can amplify 16S rRNA gene of both bacteria and archaea at the same time was used.By comparing the community changes before and after probiotics intake,it showed that this primer set is suitable for analyzing the changes of human intestinal bacteria and archaea communities.The fecal samples of volunteers were collected,and the amplification and high-throughput sequencing were carried out by using bacterial primer set(B primer)and bacterial and archaeal universal primer(AB primer);several commonly used rRNA databases were used to determine the amplification ability of the primer set to bacteria and archaea.The results showed that AB primer could display the bacterial community amplified by B primer,and could obtain the sequence of common methanogenic archaea in intestinal tract.AB primer set can analyze the bacteria and archaea in the intestinal tract at the same time by only one amplification and sequencing,which can show the structure of intestinal microbial community more comprehensively,which is suitable for the research of intestinal microorganisms.