摘要
目的探究强直性脊柱炎(ankylosing spondylitis,AS)患者肠道真菌菌群的多样性特征,分析健康人群与AS患者肠道真菌的结构差异。方法收集17例健康人群新鲜粪便样本和24例AS患者新鲜粪便样本,分别称为HC组和AS组,提取两组粪便样本总DNA;根据真菌ITS2区设计引物进行扩增,利用Illumina HiSeq PE250平台进行ITS2高通量测序;测序结果经过Reads拼接,OTUs(operational taxonomic units)聚类,Alpha和主成分分析,物种组成统计,显著性差异分析,最终得到样本物种信息。结果对肠道真菌菌群进行Alpha分析,各多样性指标中,shannon和observed_species指数差异有统计学意义(P<0.05),其余指数差异均无统计学意义,故不能明确两组间多样性的差异。主成分分析提示差异显著。对肠道真菌结构进行分析,门的水平分析显示,担子菌门(Basidiomycota)在HC组和AS组中差异显著,接合菌门(Zygomycota)在两组间差异极显著;纲和属的水平分析显示,Ascomycota_unidentified纲在两组中差异显著,伞菌纲(Agaricomycetes)、Incertae_sedis_10纲、Orbiliomycetes纲差异极显著。Agaricomycetes_unidentified_1属、Amphinema属、Geoglossales_unidentified_1属、腔块菌属(Hydnotrya)差异显著,鹅膏菌属(Amanita)和锁瑚菌属(Clavulina)差异极显著。结论本实验证实了在AS患者中存在肠道真菌菌群失调,其特征是生物多样性和结构的改变,揭示肠道真菌也可能在AS发病中发挥作用。
Objective To explore the diversity of intestinal fungi in patients with ankylosing spondylitis（AS）and analyze the differences in the structure of intestinal fungal microbiota between healthy people and AS patients.Methods The faecal specimens of 17 healthy individuals（HC group）and 24 AS patients（AS group）were collected for extraction of total DNA.The primers were designed based on fungal ITS2 region sequences,and Illumina PE250 platform was used for ITS2 high-throughput sequencing.Splicing Reads,OTUs（operational taxonomic units）clustering,species annotation,Alpha diversity analysis,Principal Coordinates analysis,species composition analysis and significant difference analysis were performed on the sequencing results to obtain the intestinal fungal information of all specimens.Results The analysis of Alpha diversity of intestinal fungal flora showed statistically significant differences in the shannon and observed_species indexes between the two groups（P〈0.05）,but not in the other diversity indexes,which means that the difference between the HC and AS groups was not clear.Principal Coordinates analysis suggested an obvious difference.The analysis on the structure of intestinal fungi showed significant difference in Basidiomycota and extremely significant difference in Zygomycota between the two groups at the phylum level.At the class and genus levels,there was significant difference in Ascomycota_unidentified between groups,while the differences in Agaricomycetes,Incertae_sedis_10 and Orbiliomycetes between the two groups were much more significant.There were significant differences in Agaricomycetes_unidentified_1,Amphinema,Geoglossales_unidentified_1 and Hydnotrya,and much more significant differences in Amanita and Clavulina between the two groups.Conclusion Our study confirmed an intestinal fungal dysbiosis characterized by alterations in biodiversity and composition in AS,suggesting that fungi might play a role in AS pathogenesis.
出处
《中国微生态学杂志》
CAS
CSCD
2018年第2期125-131,共7页
Chinese Journal of Microecology
基金
中国博士后科学基金(2016M601317)
辽宁省大学生创新创业项目(2017101610100031)
关键词
肠道真菌
强直性脊柱炎
ITS2高通量测序
Intestinal fungi
Ankylosing spondylitis
ITS2 high-throughput sequencing
