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分析微生物组成和低生物质母乳样本的共享:DNA分离和测序技术的比较
Johanne E Spreckels1, Asier Fernández-Pato1, Marloes Kruk1
1Department of Genetics, University of Groningen and University Medical Center Groningen, Groningen, the Netherlands.
ISME communications
|November 9, 2023
概括
优化研究母乳微生物组的方法至关重要. 新的DNA隔离和测序技术揭示了母亲和婴儿之间的细菌共享,即使是在低生物质样本中.
科学领域:
- 微生物学 微生物学
- 基因组学就是基因组学.
- 人类微生物组研究研究
背景情况:
- 人类牛奶微生物组研究面临的挑战是由于细菌细胞数量低和标准16SrRNA基因测序的局限性.
- 准确地描述牛奶细菌和了解母婴微生物传播对于婴儿健康至关重要.
研究的目的:
- 评估DNA分离和测序方法用于人乳微生物组分析.
- 通过优化技术,评估母婴微生物组之间的细菌共享.
主要方法:
- 测试了四种DNA分离套件 (例如DNeasy PowerSoil Pro,MagMAX),两种丰富方法和三种测序方法 (16S rRNA基因,长期读取的16S-ITS-23S rRNA基因).
- 使用模拟社区,人乳样本和方法验证的负控制.
- 分析了14对母婴对的DNA (牛奶,婴儿口服,婴儿便,母亲便).
主要成果:
- DNeasy PowerSoil Pro和MagMAX套件产生了一致的16SrRNA基因测序与低污染.
- 长期研究的16S-ITS-23S rRNA基因测序,尽管存在模拟社区偏见,但提供了可靠的牛奶样本数据,并检测了相关的母婴对之间更大的细菌共享.
- 丰富方法没有显著减少人类DNA污染.
结论:
- 使用DNeasy PowerSoil Pro或MagMAX套件与16SrRNA基因测序相结合的DNA隔离对于人类牛奶微生物群的表征是可靠的.
- 长期研究的16S-ITS-23S rRNA基因测序增强了在低生物质人乳样本中的细菌传播的研究.
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