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采用焦磷酸测序技术对无牙婴儿及其母亲或主要照顾者的唾液细菌微生物组多样性进行比较分析。

Comparative analysis of salivary bacterial microbiome diversity in edentulous infants and their mothers or primary care givers using pyrosequencing.

机构信息

Division of Nutritional Sciences, University of Illinois, Urbana, Illinois, United States of America.

出版信息

PLoS One. 2011;6(8):e23503. doi: 10.1371/journal.pone.0023503. Epub 2011 Aug 10.

Abstract

Bacterial contribution to oral disease has been studied in young children, but there is a lack of data addressing the developmental perspective in edentulous infants. Our primary objectives were to use pyrosequencing to phylogenetically characterize the salivary bacterial microbiome of edentulous infants and to make comparisons against their mothers. Saliva samples were collected from 5 edentulous infants (mean age = 4.6±1.2 mo old) and their mothers or primary care givers (mean age = 30.8±9.5 y old). Salivary DNA was extracted, used to generate DNA amplicons of the V4-V6 hypervariable region of the bacterial 16S rDNA gene, and subjected to 454-pyrosequencing. On average, over 80,000 sequences per sample were generated. High bacterial diversity was noted in the saliva of adults [1012 operational taxonomical units (OTU) at 3% divergence] and infants (578 OTU at 3% divergence). Firmicutes, Proteobacteria, Actinobacteria, and Fusobacteria were predominant bacterial phyla present in all samples. A total of 397 bacterial genera were present in our dataset. Of the 28 genera different (P<0.05) between infants and adults, 27 had a greater prevalence in adults. The exception was Streptococcus, which was the predominant genera in infant saliva (62.2% in infants vs. 20.4% in adults; P<0.05). Veillonella, Neisseria, Rothia, Haemophilus, Gemella, Granulicatella, Leptotrichia, and Fusobacterium were also predominant genera in infant samples, while Haemophilus, Neisseria, Veillonella, Fusobacterium, Oribacterium, Rothia, Treponema, and Actinomyces were predominant in adults. Our data demonstrate that although the adult saliva bacterial microbiome had a greater OTU count than infants, a rich bacterial community exists in the infant oral cavity prior to tooth eruption. Streptococcus, Veillonella, and Neisseria are the predominant bacterial genera present in infants. Further research is required to characterize the development of oral microbiota early in life and identify environmental factors that impact colonization and oral and gastrointestinal disease risk.

摘要

目前已经有研究表明,口腔疾病与细菌有关,且主要集中在幼儿群体中,但在无牙婴儿中,关于细菌对口腔疾病影响的研究还较少。本研究的主要目的是使用焦磷酸测序技术对无牙婴儿的唾液细菌微生物组进行系统发育特征分析,并与他们的母亲进行对比。收集了 5 名无牙婴儿(平均年龄 4.6±1.2 个月)及其母亲或主要照顾者(平均年龄 30.8±9.5 岁)的唾液样本。提取唾液 DNA,用于扩增细菌 16S rDNA 基因 V4-V6 高变区的 DNA 扩增子,并进行 454 焦磷酸测序。每个样本平均生成 80000 多个序列。成年人(3%差异时的 1012 个操作分类单元(OTU))和婴儿(3%差异时的 578 个 OTU)的唾液中都存在高度的细菌多样性。在所有样本中,厚壁菌门、变形菌门、放线菌门和梭杆菌门是主要的细菌门。我们的数据集中共存在 397 个细菌属。在婴儿和成人之间有 28 个不同的细菌属(P<0.05),其中 27 个在成人中更为常见。唯一的例外是链球菌,它是婴儿唾液中的主要菌属(婴儿中的 62.2%,成人中的 20.4%;P<0.05)。韦荣球菌属、奈瑟菌属、罗氏菌属、嗜血杆菌属、咽峡炎链球菌属、颗粒链菌属、勒克氏菌属和梭杆菌属也是婴儿样本中的主要菌属,而嗜血杆菌属、奈瑟菌属、韦荣球菌属、梭杆菌属、口杆菌属、罗氏菌属、密螺旋体属和放线菌属则是成人样本中的主要菌属。我们的数据表明,尽管成人唾液中的细菌微生物组 OTU 数量大于婴儿,但在出牙前,婴儿口腔中就已经存在丰富的细菌群落。链球菌属、韦荣球菌属和奈瑟菌属是婴儿口腔中主要的细菌属。需要进一步的研究来描述生命早期口腔微生物组的发育,并确定影响定植和口腔及胃肠道疾病风险的环境因素。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2ba/3154475/d51812979f9a/pone.0023503.g001.jpg

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