基于高通量测序分析孤独症谱系障碍儿童肠道菌群的构成变化

Analysis on changes of gut microbiota in children with autism spectrum disorder based on high-throughput sequencing

目的探讨孤独症谱系障碍(ASD)儿童肠道菌群的变化。方法收集35例ASD患儿(ASD组)和35名体检健康儿童(正常对照组)粪便样本,提取基因组DNA,用Illumina测序平台进行测序分析,确定群体的肠道菌群特征。结果ASD组与正常对照组的肠道菌群结构存在差异。在门水平上,ASD组与正常对照组优势菌门厚壁菌门、拟杆菌门、变形菌门及放线菌门的相对丰度差异均无统计学意义(P>0.05);糖酵母菌门在2个组之间差异有统计学意义(P=0.01),但其在肠道菌群结构中的相对丰度很低。在属水平上,ASD组与正常对照组罕见小球菌属、霍尔德曼菌属、Candidatus Saccharibacteria noname属、普雷沃菌属、Burkholderiales noname属、戈登杆菌属、另枝菌属、Solobacterium属、Parasutterella属、Anaerotruncus属、杆菌属、丛毛单胞菌属、孪生球菌属及颗粒球菌属的相对丰度差异有统计学意义(P<0.05)。进一步对菌群进行分析,ASD组肠道菌群中翁德顿氏另枝菌、Alistipes putredinis、芬戈尔德氏拟杆菌、谢氏另枝菌、前庭链球菌、丝状霍尔德曼菌、芽胞拟杆菌、伯克霍尔德菌-1-1-47、单形巨单胞菌、粪副拟杆菌、生黄瘤胃球菌、毛螺科菌_3_1_57FAA_CT1及Parasutterella excrementihominis的相对丰度均高于正常对照组,毗邻颗粒链球菌、死亡梭杆菌、莫氏细小杆菌、直肠真杆菌、Candidate division_TM7_single_cell_isolate_TM7c及麻疹孪生球菌相对丰度均低于正常对照组(P<0.05)。结论ASD患儿存在肠道微生态失调,但无特定的肠道微生物群组成。

Objective To investigate the changes of gut microbiota in children with autism spectrum disorders(ASD).Methods A total of 35 children with ASD were enrolled into ASD group,and 35 healthy children were assigned to healthy control group.Genomic DNA was extracted from fecal samples,and Illumina sequencing platform was used for sequencing analysis to determine the characteristics of gut microbiota.Results There was difference in gut microbiota between ASD and healthy control groups.At the phylum level,there was no significant difference in the relative abundance of the Firmicutes,Bacteroidetes,Proteobacteria or Actinobacteria(P>0.05).There was a significant difference in the structure of Candidatus Saccharibacteria between the 2 groups(P=0.01),but the relative abundance in the structure of gut microbiota was very low.At the genus level,there were significant differences in the relative abundance of Subdoligranulum,Holdemania,Candidatus Saccharibacteria noname,Prevotella,Burkholderiales noname,Gordonibacter,Alistipes,Solobacterium,Parasutterella,Anaerotruncus,Aggregatibacter,Comamonas,Gemella and Granulicatella between the 2 groups(P<0.05).Further analysis of gut microbiota showed that the relative abundance of Alistipes onderdonkii,Alistipes putredinis,Bacteroides finegoldii,Alistipes shahii,Streptococcus vestibularis,Holdemania filiformis,Bacteroides massiliensis,Burkholderiales bacterium-1-1-47,Megamonas funiformis,Parabacteroides merdae,Ruminococcus flavefaciens,Lachnospiraceae bacterium_3_1_57FAA_CT1 and Parasutterella excrementihominis in ASD group were higher than those in healthy control group,but the relative abundance of Granulicatella adiacens,Fusobacterium mortiferum,Solobacterium moorei,Eubacterium rectale,Candidate division_TM7_single_cell_isolate_TM7c and Gemella morbillorum in the ASD froup were lower than those in the healthy control group(P<0.05).Conclusion Intestinal microecological disorders exist in ASD children,but ASD children do not have a specific intestinal microbial composition.