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目的探讨克罗恩病(CD)患者肠道菌群多样性变化。方法收集8例确诊CD患者的粪便标本(CD组),以同期8例健康人群粪便作为对照组(CN组)。收集新鲜粪便样本提取DNA,并将样本进行PCR扩增,最终进行16SrRNA数据分析。结果测序共获得990 890条高质量序列,CD组和CN组多样性指数Shannon和Simpson比较差异有统计学意义(t=-3.802和t=-2.886,P<0.05)。OTUs(可操作分类学单位)比较韦恩图显示CD组共有7 437株菌种,而CN组共有7 744株菌种,其中3 438株菌种重叠。LDA差异贡献分析图提示CD患者肠道菌群以芽胞杆菌、变形菌、乳杆菌目、放线菌目、异常球菌纲、栖热菌目、假单胞菌目和交替单胞菌目等为主要特征差异细菌菌类,而CN组则以双歧杆菌目及双歧杆菌科,Odoribacteraceae、Christensenellaceae、弧菌目及弧菌科等为主要特征差异细菌菌类。据物种进化树的样本菌落分布图显示:两组样本厚壁菌门、放线菌门、拟杆菌门、变形菌门、疣微菌门、酸杆菌门和螺旋菌门占主要门类分布。CD组及CN组细菌科类分析结果:毛螺菌科、韦荣球菌科、瘤胃菌科、丹毒丝菌科、红蝽菌科、紫单胞菌科、双歧杆菌科、普雷沃菌科、拟杆菌科、产碱菌科和链球菌科在两组样本中共同存在。同时从该进化树可发现,CD组中红蝽菌科、产碱菌科、丹毒丝菌科和链球菌科相对于CN组优势丰度分布,而普雷沃菌科、拟杆菌科和双歧杆菌科比CN组丰度明显减少。此外,Dethiosulfovibrionaceae、丙酸杆菌科、盐单胞菌科、希万菌科等为CD组特有菌种。结论 CD组的菌群多样性明显少于CN组,CD组有其特有优势菌群。
Objective To investigate the changes of intestinal microflora in patients with Crohn’s disease (CD). Methods Stool samples from 8 patients diagnosed with CD (CD group) were collected. The faeces of 8 healthy people in the same period were taken as the control group (CN group). Fresh stool samples were collected for DNA extraction, and samples were PCR-amplified for 16S rRNA data analysis. Results A total of 990 890 high-quality sequences were obtained by sequencing. The Shannon and Simpson indices of diversity in CD group and CN group were significantly different (t = -3.802 and t = -2.886, P <0.05). OTUs (operable taxa units) Comparison of Wayne diagram showed a total of 7 437 strains of CD strains, CN group, a total of 7 744 strains, of which 3 438 strains overlap. LDA differential contribution analysis showed that the intestinal flora of patients with Bacillus Bacillus, Proteus, Lactobacillus, Actinomycetes, Anomalae, Thermus, Pseudomonas and Alternaria The main characteristics of different bacterial flora, while the CN group Bifidobacteria and Bifidobacteria, Odoribacteraceae, Christensenellaceae, Vibrio and Vibrio families as the main characteristics of different bacterial flora. According to the species colonies distribution map of the sample colonies showed: two groups of specimens of Firmicutes, actinomycetes, Bacteroidetes, Proteobacteria, warts, mycobacteria and spirochetes accounted for the main categories. The results of the bacterial families in CD group and CN group were as follows: Mycoplasma, Veillonella, Rumenaceae, Erysipelothrix, Taxobinidae, Pseudomonas, Bifidobacteria, Prevotella Section, Bacteroidetes, Alcaligenes and Streptococci co-exist in both sets of samples. At the same time, it can be found from this phylogenetic tree that the distributions of the dominant taxonomic species of Taxobin, Alcaligenes, Erysipelas and Streptococci in the CD group are relative to those in the CN group. However, Bifidobacterium Bryant CN group significantly reduced abundance. In addition, Dethiosulfovibrionaceae, Propionibacteriaceae, Salmonellaceae, Xegenomycetes, etc. are endemic strains of CD group. Conclusion The bacterial diversity of CD group was significantly less than that of CN group, and CD group had its own dominant microflora.