基于16S rRNA测序比较探讨气虚证患者与健康人的肠道菌群的结构特征＊

目的 采用16S rRNA测序技术,检测比较气虚患者和健康人群的肠道菌群多样性、相对丰度及分布的差异。方法 选取在湖北省中医院就诊的气虚证患者20例为观察对象,以及同期健康志愿者10名作为对照。采用粪便基因组DNA提取试剂盒抽提HG和QD粪便样本中DNA,对V3-V4区进行PCR扩增,再进行文库构建和Illumina MiSeq测序。最后对数据进行生物信息学分析,探索气虚证人群肠道菌群的结构组成特征。结果 本研究共得到534个OTU。物种累积曲线表明本研究所收集的样本量充足。HG和QD两组alpha指数不存在明显差异（P＞0.05）。通过beta多样性分析可见,HG和QD两组肠道菌群构成存在一定差异。此外,QD组的Firmicutes/Bacteroides比值较健康人群升高。相较于HG,g_Odoribacter、蓝藻菌门、g_Anaerotruncus、c_Chloroplast、s_Clostridium_Methylpentosum和o_Streptophyta在QD中显著富集（P < 0.05）。结论 气虚证患者肠道菌群的组成结构与健康人群存在一定差异。在气虚证的发生发展中,肠道菌群的改变可能起到了重要作用。

Comparing the Structural Characteristics of Gut Microbiota in Patients with Qi Deficiency Syndrome and Healthy Individuals Based on 16S rRNA Sequencing

Objective To compare the diversity, relative abundance, and distribution of intestinal flora between Qi deficiency patients and healthy people by 16S rRNA sequencing.Methods 20 patients with Qi deficiency syndrome in Hubei Hospital of Traditional Chinese Medicine were selected as the observation objects, and 10 healthy volunteers were selected as the control group. A genomic DNA extraction kit was used to extract DNA from HG and QD fecal samples. PCR amplification of the V3-V4 region was applied, and library construction and Illumina Miseq sequencing were performed. Finally, bioinformatics analysis of the data was carried out to explore the structural characteristics of the intestinal flora of the Qi deficiency group.Results A total of 534 OTUs were obtained. The species accumulation curve showed that the sample size was sufficient. There was no significant difference in the alpha diversity between HG and QD (P > 0.05). According to the beta diversity analysis, the composition of intestinal flora differed to some extent between HG and QD. Besides, the Firmicutes / Bacteroides ratio was elevated in the QD group compared with the healthy population. Compared with HG, g_Odoribacter, p_cyanobacteria, g_Anaerotruncus, c_Chloroplast, s_Clostridium_Methylpentosum, and o_Streptophyta was remarkably increased in QD (P < 0.05).Conclusion The compositional structure of gut microbiota in patients diagnosed with Qi deficiency differs from that in healthy individuals. The change of intestinal flora may play an important role in the occurrence and development of Qi deficiency syndrome.