孤独症谱系障碍儿童的唾液微生物群落研究

Study of salivary microbiome in children with autism spectrum disorders

目的:通过宏基因组学方法结合Pacbio SMRT高通量测序技术探究孤独症谱系障碍(ASD)儿童中口腔健康和罹患龋齿者的唾液微生物菌群的组成及其差异。方法:收集42名ASD儿童唾液样本,其中无龋组22名,高龋组20名,提取DNA进行聚合酶链式反应扩增,利用PacBio SMRT高通量测序技术对16S rRNA全长进行测序,进行口腔菌群结构和组成的分析比较。结果:两组间唾液菌群结构差异有统计学意义(P<0.05),且高龋组的菌落结构更为相似和保守(P<0.05)。在种水平上,无龋组高表达有伯杰氏杆菌、南锡普氏菌、金黄色葡萄球菌三个健康相关微生物(P<0.05),高龋组高表达有变形链球菌、口普氏菌、浑浊戴阿利斯特杆菌、具核梭杆菌四个龋病相关微生物(P<0.05)。结论:ASD儿童的龋齿唾液菌群有整体菌群结构和特定物种组成的改变,为ASD儿童的龋齿预防和诊断提供新策略。

Objective: The salivary microbial communities in healthy and caries children with autism spectrum disorders(ASD) were compared by third high-throughput sequencing platform combined with metagenomics technique.Methods: Saliva samples were collected from 22 healthy children(Caries-free, CF group) and 20 high caries(Caries-high, CH group) children with ASD, and the metagenomic DNA was extracted and amplified by polymerase chain reaction(PCR). The full-length 16S rRNA gene was sequenced by PacBio SMRT sequencing technique, and the difference of salivary microbial structure and composition was analyzed and compared between the two groups.Results: The difference in microbial structure between the CF and CH group was statistically significant(P<0.05). Moreover, the community structure in CH group was more similar and conserved(P<0.05). The microbial composition analysis results identified three health-enriched taxa, namely,Bergeyella spp,Prevotella nanceiensis, andStaphylococcus aureusfor CF group(P<0.05), and four caries-enriched taxa includingStreptococcus mutans,Prevotella oris,Dialister invisusandFusobacterium nucleatum for CH group at species level(P<0.05).Conclusions: The study characterized the caries-specific community structure and biomarkers of the oral microbiota in ASD children, which might help develop novel strategies for the prevention and diagnosis of dental caries in ASD children population.