基于二代测序的肝泡型包虫病患者肠道菌群差异性分析  

Differential analysis of intestinal flora in patients with hepatic blastomycosis based on second-generation sequencing

作　　者：马小亚 阿祥仁[2] 马继登 周健武 程鹏 唐赟[1] Ma Xiaoya;A Xiangren;Ma Jideng;Zhou Jianwu;Cheng Peng;Tang Yun(Department of Basic Medicine,College of Medicine,Qinghai University,Xining 810016,China;Department of Medical Laboratory,Qinghai Provincial People′s Hospital,Xining 810007,China)

机构地区：[1]青海大学医学院基础医学部,西宁810016 [2]青海省人民医院医学检验科,西宁810007

出　　处：《中华预防医学杂志》2025年第1期101-109,共9页Chinese Journal of Preventive Medicine

基　　金：青海省2022年创新平台建设专项项目(2022-ZJ-T01)。

摘　　要：本研究通过宏基因组测序技术,探讨肝泡型包虫患者肠道菌群的差异性,寻找疾病进程中可能起作用的肠道微生物成员。采用病例对照研究,纳入2023年10月至2024年1月期间,青海省人民医院收治的肝泡型包虫病患者及就诊健康体检的个体的粪便样本。包括实验组(AE组)10例肝泡型包虫病的患者,对照组(NC组)9例健康体检者。采用Illumina Novaseq 6000测序平台对这两组样本进行宏基因组测序,使用fastp(v0.20.1)去除接头,bbmap(v38.93-0)去除宿主化序列,MEGAHIT(v1.2.9)进行序列拼接,prodigal(v2.6.3)对拼接的scaffold进行ORF预测并翻译为氨基酸序列,MMSeqs2(v13.45111)构建非冗余基因集,salmon(v1.8.0)与非冗余基因集进行比对。通过非冗余数据库对物种进行注释,计算出各样本中的物种丰度,用Wilcoxon秩和检验对两组进行检验。最后利用线性判别分析统计两组肠道菌群差异,利用冗余分析(RDA)分析肠道差异菌群与临床指标之间的相关性。结果显示,各样本的有效数据量分布在10.41~12.46 G,去冗余后构建gene catalogue(非冗余基因集)中ORF数目为4951408,非冗余基因与NR数据库比对,其注释率为97.97%。两组研究对象年龄分别为:NC组(44.78±4.58)岁,AE组(42.90±10.44)岁,差异无统计学意义(t=0.530,P=0.476)。两组体重指数(BMI)(t=2.368,P=0.142)、性别(χ^(2)=0.200,P=0.655)、饮食习惯相匹配。AE组α多样性没有显著的统计学差异(ACE指数,t=0.942;chao1指数,t=0.947;shannon指数,t=0.813,simpson指数,t=0.613,P>0.05),而β多样性分析显示两组群落的整体结构上存在明显的差异(Stress=0.0545)。在门水平一共注释到120个物种,其中有差异的有2个。而属水平注释到1736个物种,有差异的为69个,在种水平有差异的是309个。AE组属水平上拟杆菌属、埃希氏菌属、梭菌属、另枝菌属、瘤胃球菌属、颤螺菌属丰度排名前6;而NC组中西加特罗拉属(Segatella)、普雷沃氏菌属、粪Exploring the variability of the intestinal flora of patients with hepatic blastocysticercosis and searching for members of the intestinal microflora that may play a role in the disease process by means of macro-genome sequencing technology.A case-control study was used to include fecal samples from patients with hepatic vesicular schistosomiasis admitted to Qinghai Provincial People′s Hospital between October 2023 and January 2024 and individuals attending health checkups.The experimental group(AE group)consisted of 10 patients with liver vesicular schistosomiasis and the control group(NC group)consisted of 9 individuals attending health checkups.Macrogenomic sequencing was performed on these two groups of samples using the Illumina Novaseq 6000 sequencing platform,using fastp(v0.20.1)to remove junctions,and bbmap(v38.93-0)to remove the hosted sequences,followed by sequence splicing using MEGAHIT(v1.2.9),and then using prodigal(v2.6.3)to The spliced scaffold was subjected to ORF prediction and translated into amino acid sequences,followed by the construction of a non-redundant gene set using MMSeqs2(v13.45111),and finally compared with the non-redundant gene set using salmon(v1.8.0).Species were annotated by the non-redundant database,species abundance was calculated in each sample,and the two sets were tested using Wilcoxon rank sum test.Finally,the differences in intestinal flora between the two groups were statistically analyzed using linear discriminant analysis,and the correlation between the differential intestinal flora and clinical indicators was analyzed using redundancy analysis(RDA).The results showed that the effective data volume of each sample was distributed from 10.41 to 12.46 G.The number of ORFs in the de-redundantly constructed gene catalogue(non-redundant gene set)was 4951408,and the annotation rate of the non-redundant genes was 97.97%when compared with the NR database.The ages of the study subjects in the two groups were(44.78±4.58)years in the NC group and(42.90±10.44)years in the AE g

关 键 词：肝泡型包虫病 肠道菌群 差异性 宏基因组测序 

分 类 号：R532.32[医药卫生—内科学]