使用流细胞计量量化合成细菌群体组成:在模拟群体中的有效性和共同培养中的挑战
Fabian Mermans1,2, Ioanna Chatzigiannidou1, Wim Teughels2
1Ghent University, Center for Microbial Ecology and Technology (CMET), Faculty of Bioscience Engineering, Gent, Belgium.
mSystems
|November 29, 2024
概括
流细胞计为量化细菌群体提供了一种快速,廉价的方法. 然而,由于表型变化,它的准确性在共同培养下降,这给微生物研究带来了挑战.
科学领域:
- 微生物学 微生物学
- 生物技术是生物技术.
- 生物信息学是一种生物信息学.
背景情况:
- 准确的细菌群体组成确定对于微生物系统研究至关重要.
- 传统的方法,如和PCR是劳动密集型,昂贵,容易产生偏见.
- 流细胞计是社区分析的有希望,快速和经济有效的替代方案.
研究的目的:
- 评估流细胞计的性能,以量化口服合成细菌群落.
- 为了比较流细胞计的准确性与菌株特异性定量PCR (qPCR) 和16S rRNA基因安普利康序列.
- 评估共同培养对流细胞计在确定社区组成的准确性的影响.
主要方法:
- 准备的菌培养,模拟社区和口腔细菌的共同培养.
- 使用随机森林分类器,这些分类器是根据来自轴基培养的流动细胞计数据进行训练的.
- 使用流细胞计,菌株特异性qPCR和16S rRNA基因安普利康序列的量化社区组成.
主要成果:
- 流细胞测量显示了较低的平均根平均平方误差,在模拟社区中表现优于PCR方法.
- 流细胞计和基于PCR的方法都没有准确地反映共同种群中的社区组成.
- 由于细菌表型的改变,共培养物中流细胞计性能下降,与qPCR相比,发现了差异.
结论:
- 流细胞计显示了量化细菌群落的潜力,特别是在更简单的模拟群落中.
- 联合培养由于表型可塑性而显著影响流动细胞计的准确性.
- 需要进一步的研究来克服应用流细胞计对复杂的共同培养微生物群落的挑战.
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