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基于高度准确的 16S rRNA 基因测序分析的儿童鼻分泌物中的细菌组成。

Bacterial composition of nasal discharge in children based on highly accurate 16S rRNA gene sequencing analysis.

机构信息

Department of Microbiology, School of Medicine, University of Occupational and Environmental Health, Japan, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu, 807-8555, Japan.

Department of Pediatrics, School of Medicine, University of Occupational and Environmental Health, Japan, Kitakyushu, Japan.

出版信息

Sci Rep. 2020 Nov 19;10(1):20193. doi: 10.1038/s41598-020-77271-z.

Abstract

Nasopharyngeal colonization by bacteria is a prerequisite for progression to respiratory disease and an important source of horizontal spread within communities. We aimed to perform quantitative analysis of the bacterial cells and reveal the microbiota of the nasal discharge in children at the species level based on highly accurate 16S rRNA gene sequencing. This study enrolled 40 pediatric patients with rhinorrhea. The bacterial cells in the nasal discharge were counted by epifluorescence microscopic analysis. The microbiota was analyzed by using the 16S rRNA gene clone library sequencing method. We demonstrated that a high abundance (median 2.2 × 10 cells/mL) of bacteria was contained in the nasal discharge of children. Of the 40 samples, 37 (92.5%) were dominated by OTUs corresponding to Haemophilus aegyptius/influenzae, Moraxella catarrhalis/nonliquefaciens, or Streptococcus pneumoniae. These samples showed higher cell abundance and lower alpha diversity than the remaining three samples in which the other bacteria coexisted. In addition, 12 sequences with low homology to type strains were considered as previously unknown bacterial lineages. In conclusion, the nasal discharge of most young children contains a large amount of respiratory pathogens and several unknown bacteria, which could not only cause endogenous infection but also be a source of transmission to others.

摘要

鼻咽部细菌定植是引发呼吸道疾病的前提条件,也是社区内水平传播的重要来源。本研究旨在通过高精确性 16S rRNA 基因测序对儿童鼻分泌物进行定量分析,并在种属水平上揭示其微生物群。本研究纳入了 40 例有流涕症状的儿科患者。采用荧光显微镜分析技术对鼻分泌物中的细菌细胞进行计数,并用 16S rRNA 基因克隆文库测序方法对微生物群进行分析。结果表明,儿童鼻分泌物中含有大量细菌(中位数 2.2×10 细胞/ml)。在 40 个样本中,37 个(92.5%)样本以 Haemophilus aegyptius/influenzae、Moraxella catarrhalis/nonliquefaciens 或 Streptococcus pneumoniae 的 OTUs 为主。这些样本的细胞丰度较高,α多样性较低,而其余三个共存其他细菌的样本则较低。此外,12 个与模式株同源性较低的序列被认为是以前未知的细菌谱系。综上所述,大多数幼儿的鼻分泌物中含有大量的呼吸道病原体和几种未知细菌,这些细菌不仅会引发内源性感染,还可能成为向他人传播的源头。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b26a/7678852/ad29c7393551/41598_2020_77271_Fig1_HTML.jpg

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