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

doi:10.3969/j.issn.1673-8640.2021.02.007

摘要/Abstract

摘要：

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。

关键词: 肠道菌群, 高通量测序, 孤独症谱系障碍

Abstract:

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.

Key words: Gut microbiota, High-throughput sequencing, Autism spectrum disorder

中图分类号:

R372

曾佩佩, 曾婷, 邓梁琼, 郭浩, 冯玉山, 黄丽萍, 李红辉. 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[J]. 检验医学, 2021, 36(2): 153-161.

ZENG Peipei, ZENG Ting, DENG Liangqiong, GUO Hao, FENG Yushan, HUANG Liping, LI Honghui. Analysis on changes of gut microbiota in children with autism spectrum disorder based on high-throughput sequencing[J]. Laboratory Medicine, 2021, 36(2): 153-161.

图/表 10

参考文献 24

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组别	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
ASD组	2.08	0.04	0.002	3.61	0.09	0.012	6.34
正常对照组	0.74	0.02	0.009	3.39	0.04	0.01	2.44
统计值	842.0	827.0	471.0	814.0	802.0	800.0	808.0
P值	0.007	0.008	0.017	0.018	0.018	0.02	0.022
组别	Solobacterium属	Parasutterella属	Anaerotruncus属	杆菌属	丛毛单胞菌属	孪生球菌属	颗粒球菌属
ASD组	0.001	0.14	0.16	0	0.001	0.012	0.012
正常对照组	0.002	0.05	0.09	0.001	0	0.03	0.04
统计值	469.5	793.0	785.0	542.5	682.5	444.0	444.0
P值	0.022	0.03	0.042	0.042	0.042	0.046	0.048

组别	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
ASD组	2.08	0.04	0.002	3.61	0.09	0.012	6.34
正常对照组	0.74	0.02	0.009	3.39	0.04	0.01	2.44
统计值	842.0	827.0	471.0	814.0	802.0	800.0	808.0
P值	0.007	0.008	0.017	0.018	0.018	0.02	0.022
组别	Solobacterium属	Parasutterella属	Anaerotruncus属	杆菌属	丛毛单胞菌属	孪生球菌属	颗粒球菌属
ASD组	0.001	0.14	0.16	0	0.001	0.012	0.012
正常对照组	0.002	0.05	0.09	0.001	0	0.03	0.04
统计值	469.5	793.0	785.0	542.5	682.5	444.0	444.0
P值	0.022	0.03	0.042	0.042	0.042	0.046	0.048

组别	翁德顿氏另枝菌		Alistipes putredinis		毗邻颗粒链球菌		芬戈尔德氏拟杆菌	谢氏另枝菌	前庭链球菌	死亡梭杆菌	丝状霍尔德曼菌	芽胞拟杆菌	伯克霍尔德菌-1-1-47
ASD组	1.636		3.699		0.002		0.290	0.679	0.025	0.032	0.028	1.665	0.095
正常对照组	0.150		1.686		0.006		0.037	0.187	0.004	0.557	0.007	0.681	0.037
统计值	915.0		849.0		415.0		811.0	821.0	784.0	452.0	786.5	814.0	802.0
P值	0.000 2		0.005		0.008		0.009	0.011	0.013	0.014	0.016	0.017	0.018
组别	单形巨单胞菌	Gordonibacter pamelaeae	粪副拟杆菌	生黄瘤胃球菌		莫氏细小杆菌	毛螺科菌_3_1_ 57FAA_CT1	Parasutterella excrementihominis		直肠真杆菌	Candidate division_TM7_single_cell_isolate_TM7c		麻疹孪生球菌
ASD组	0.212	0.018	1.068	0.008 0		0.001	0.045	0.139		2.549	0.001		0.000 3
正常对照组	0.120	0.007	0.886	0.000 3		0.002	0.011	0.049		5.018	0.006		0.004 0
统计值	785.0	800.0	807.5	737.0		469.5	780.5	793.0		794.0	488.0		488.0
P值	0.019	0.020	0.020	0.021		0.023	0.026	0.030		0.032	0.032		0.032

组别	翁德顿氏另枝菌		Alistipes putredinis		毗邻颗粒链球菌		芬戈尔德氏拟杆菌	谢氏另枝菌	前庭链球菌	死亡梭杆菌	丝状霍尔德曼菌	芽胞拟杆菌	伯克霍尔德菌-1-1-47
ASD组	1.636		3.699		0.002		0.290	0.679	0.025	0.032	0.028	1.665	0.095
正常对照组	0.150		1.686		0.006		0.037	0.187	0.004	0.557	0.007	0.681	0.037
统计值	915.0		849.0		415.0		811.0	821.0	784.0	452.0	786.5	814.0	802.0
P值	0.000 2		0.005		0.008		0.009	0.011	0.013	0.014	0.016	0.017	0.018
组别	单形巨单胞菌	Gordonibacter pamelaeae	粪副拟杆菌	生黄瘤胃球菌		莫氏细小杆菌	毛螺科菌_3_1_ 57FAA_CT1	Parasutterella excrementihominis		直肠真杆菌	Candidate division_TM7_single_cell_isolate_TM7c		麻疹孪生球菌
ASD组	0.212	0.018	1.068	0.008 0		0.001	0.045	0.139		2.549	0.001		0.000 3
正常对照组	0.120	0.007	0.886	0.000 3		0.002	0.011	0.049		5.018	0.006		0.004 0
统计值	785.0	800.0	807.5	737.0		469.5	780.5	793.0		794.0	488.0		488.0
P值	0.019	0.020	0.020	0.021		0.023	0.026	0.030		0.032	0.032		0.032

组别	K08856	K02319	K04464	K06322	K09385	K09628	K09655	K12444	K12688	K13003
ASD组	9.257	90.714	20.571	50.829	22.600	15.257	38.200	78.343	4.429	24.514
正常对照组	109.657	1.000	7.457	4.571	1.771	5.257	7.543	11.143	62.257	2.257
P值	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00
组别	K14446	K14819	K16323	K17916	K18563	K18663	K19562	K20891	K21285	K21559
ASD组	21.429	46.457	23.543	30.200	21.829	1.971	1.143	23.886	35.800	60.286
正常对照组	4.629	4.171	4.371	91.114	1.600	17.743	178.400	1.343	2.743	17.829
P值	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00	0.00